{ "cells": [ { "cell_type": "markdown", "metadata": {}, "source": [ "# Analysing simulated and empirical datasets with stacks, pyrad, and ipyrad\n", "\n", "This ipython notebook will provide a completely reproducible record of\n", "all the analyses for the ipyrad manuscript.\n", "\n", "All analyses were performed on a 40 core Intel(R) Xeon(R) CPU E5-2680 v2 @ 2.80GHz \n", "Silicon Mechanics compute node with 120GB of main memory running CentOS Linux release 7.1.1503.\n", "\n", "* Insert thanks to the CCNY scientific computing cluster folks here\n", "\n", "\n", "### Notebook Tunnels\n", "Mostly notes for myself, since my cluster config is complicated. To get a connection to a notebook on a compute node I have to bounce of 2 different machines, a gateway to get on campus, then another gateway (head hpc node) so I can talk to the cluster. The ssh commands will spawn processes in the background to shuttle communication between the ports. Kinda fancy.\n", "\n", "On the cluster compute node (where you actually want to do the work) run:\n", "\n", " ipython notebook --no-browser --port=8888 &\n", " \n", "On the cluster head node run this to forward connections from the campus to a node inside the hpc infrastructure:\n", "\n", " ssh -N -f -L localhost:8888:localhost:8888 \n", "\n", "On your gateway box run:\n", "\n", " ssh -N -f -L localhost:8887:localhost:8888 @\n", "\n", "\n", "## Prerequisites (download and install data and executables)" ] }, { "cell_type": "code", "execution_count": 1, "metadata": { "collapsed": false }, "outputs": [], "source": [ "import shutil\n", "import glob\n", "import sys\n", "import os\n", "\n", "## Set the default directories for exec and data. \n", "WORK_DIR=\"/home/iovercast/manuscript-analysis/\"\n", "EMPERICAL_DATA_DIR=os.path.join(WORK_DIR, \"example_empirical_rad/\")\n", "SIMULATION_DATA_DIR=os.path.join(WORK_DIR, \"simulated_data\")\n", "IPYRAD_DIR=os.path.join(WORK_DIR, \"ipyrad/\")\n", "PYRAD_DIR=os.path.join(WORK_DIR, \"pyrad/\")\n", "STACKS_DIR=os.path.join(WORK_DIR, \"stacks/\")\n", "AFTRRAD_DIR=os.path.join(WORK_DIR, \"aftrRAD/\")\n", "DDOCENT_DIR=os.path.join(WORK_DIR, \"dDocent/\")\n", "\n", "## (emprical data dir will be created for us when we untar it)\n", "for dir in [WORK_DIR, IPYRAD_DIR, PYRAD_DIR, STACKS_DIR, AFTRRAD_DIR, DDOCENT_DIR]:\n", " if not os.path.exists(dir):\n", " os.makedirs(dir)\n", "\n", "## Empirical output directories\n", "IPYRAD_OUTPUT=os.path.join(IPYRAD_DIR, \"REALDATA/\")\n", "PYRAD_OUTPUT=os.path.join(PYRAD_DIR, \"REALDATA/\")\n", "STACKS_OUTPUT=os.path.join(STACKS_DIR, \"REALDATA/\")\n", "STACKS_GAP_OUT=os.path.join(STACKS_OUTPUT, \"gapped/\")\n", "STACKS_UNGAP_OUT=os.path.join(STACKS_OUTPUT, \"ungapped/\")\n", "STACKS_DEFAULT_OUT=os.path.join(STACKS_OUTPUT, \"default/\")\n", "AFTRRAD_OUTPUT=os.path.join(AFTRRAD_DIR, \"REALDATA\")\n", "DDOCENT_OUTPUT=os.path.join(DDOCENT_DIR, \"REALDATA\")\n", "\n", "## Make the empirical output directories if they don't already exist\n", "for dir in [IPYRAD_OUTPUT, PYRAD_OUTPUT, STACKS_OUTPUT,\\\n", " STACKS_GAP_OUT, STACKS_UNGAP_OUT, STACKS_DEFAULT_OUT, AFTRRAD_OUTPUT, DDOCENT_OUTPUT]:\n", " if not os.path.exists(dir):\n", " os.makedirs(dir)\n", "\n", "os.chdir(WORK_DIR)\n" ] }, { "cell_type": "code", "execution_count": 4, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ " % Total % Received % Xferd Average Speed Time Time Time Current\n", " Dload Upload Total Spent Left Speed\n", "100 1042M 100 1042M 0 0 17.8M 0 0:00:58 0:00:58 --:--:-- 14.5M\n", "example_empirical_rad/\n", "example_empirical_rad/38362_rex.fastq.gz\n", "example_empirical_rad/32082_przewalskii.fastq.gz\n", "example_empirical_rad/40578_rex.fastq.gz\n", "example_empirical_rad/30686_cyathophylla.fastq.gz\n", "example_empirical_rad/39618_rex.fastq.gz\n", "example_empirical_rad/41954_cyathophylloides.fastq.gz\n", "example_empirical_rad/41478_cyathophylloides.fastq.gz\n", "example_empirical_rad/33588_przewalskii.fastq.gz\n", "example_empirical_rad/35855_rex.fastq.gz\n", "example_empirical_rad/35236_rex.fastq.gz\n", "example_empirical_rad/29154_superba.fastq.gz\n", "example_empirical_rad/30556_thamno.fastq.gz\n", "example_empirical_rad/33413_thamno.fastq.gz\n" ] } ], "source": [ "### Fetch the pedicularis data\n", "\n", "##curl grabs the data from a public dropbox url\n", "## the curl command uses an upper-case o argument, not a zero.\n", "!curl -LkO https://dl.dropboxusercontent.com/u/2538935/example_empirical_rad.tar.gz\n", "\n", "## the tar command decompresses the data directory\n", "!tar -xvzf example_empirical_rad.tar.gz\n" ] }, { "cell_type": "code", "execution_count": 195, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ " % Total % Received % Xferd Average Speed Time Time Time Current\n", " Dload Upload Total Spent Left Speed\n", "100 93.6M 100 93.6M 0 0 12.1M 0 0:00:07 0:00:07 --:--:-- 14.8M\n", "./._Hmel2\n", "Hmel2/\n", "Hmel2/._annotation\n", "Hmel2/annotation/\n", "Hmel2/._ChangeLog.txt\n", "Hmel2/ChangeLog.txt\n", "Hmel2/._Hmel2.fa\n", "Hmel2/Hmel2.fa\n", "Hmel2/._Hmel_mtDNA.fa\n", "Hmel2/Hmel_mtDNA.fa\n", "Hmel2/._maps\n", "Hmel2/maps/\n", "Hmel2/._README.txt\n", "Hmel2/README.txt\n", "Hmel2/._repeats\n", "Hmel2/repeats/\n", "Hmel2/._transfer\n", "Hmel2/transfer/\n", "Hmel2/transfer/._Hmel1-1_Hmel2.chain\n", "Hmel2/transfer/Hmel1-1_Hmel2.chain\n", "Hmel2/transfer/._Hmel2_broken.gff\n", "Hmel2/transfer/Hmel2_broken.gff\n", "Hmel2/transfer/._Hmel2_removed.gff\n", "Hmel2/transfer/Hmel2_removed.gff\n", "Hmel2/transfer/._Hmel2_transfer_new.tsv\n", "Hmel2/transfer/Hmel2_transfer_new.tsv\n", "Hmel2/transfer/._Hmel2_transfer_old.tsv\n", "Hmel2/transfer/Hmel2_transfer_old.tsv\n", "Hmel2/repeats/._Hmel.all.named.final.1-31.lib\n", "Hmel2/repeats/Hmel.all.named.final.1-31.lib\n", "Hmel2/maps/._Hmel2.gaps.bed\n", "Hmel2/maps/Hmel2.gaps.bed\n", "Hmel2/maps/._Hmel2_chromosome_linkage.tsv\n", "Hmel2/maps/Hmel2_chromosome_linkage.tsv\n", "Hmel2/maps/._Hmel2_chromosomes.agp\n", "Hmel2/maps/Hmel2_chromosomes.agp\n", "Hmel2/maps/._Hmel2_scaffold_linkage.tsv\n", "Hmel2/maps/Hmel2_scaffold_linkage.tsv\n", "Hmel2/maps/._Hmel2_scaffolds.agp\n", "Hmel2/maps/Hmel2_scaffolds.agp\n", "Hmel2/annotation/._Hmel2.gff\n", "Hmel2/annotation/Hmel2.gff\n", "Hmel2/annotation/._Hmel2_cds.fa\n", "Hmel2/annotation/Hmel2_cds.fa\n", "Hmel2/annotation/._Hmel2_proteins.fa\n", "Hmel2/annotation/Hmel2_proteins.fa\n", "Hmel2/annotation/._Hmel_mtDNA.gff\n", "Hmel2/annotation/Hmel_mtDNA.gff\n" ] } ], "source": [ "## The original Helocnius analysis from pyrad v3\n", "## Fetch the heliconius genome and rad data\n", "##\n", "## Davey, John W., et al. \"Major improvements to the Heliconius melpomene \n", "## genome assembly used to confirm 10 chromosome fusion events in 6 \n", "## million years of butterfly evolution.\" G3: Genes| Genomes| Genetics 6.3\n", "## (2016): 695-708.\n", "\n", "!curl -LkO http://butterflygenome.org/sites/default/files/Hmel2-0_Release_20160201.tgz\n", "!tar -xvzf Hmel2-0_Release_20160201.tgz\n", "\n", "## Several RAD datasets are available\n", "##\n", "## Heliconius Genome Consortium. (2012). Butterfly genome reveals \n", "## promiscuous exchange of mimicry adaptations among species. \n", "## Nature, 487(7405), 94-98.\n", "## European Nucleotide Archive, Accession ERP000991\n", "## Heliconius melpomene melpomene x Heliconius melpomene rosina - ERP000993\n", "\n", "## And from the Davey et al 2016\n", "## European Nucleotide Archive (ENA), accession PRJEB11288" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## Prereqs - Install ipyrad\n", "\n", "Full install details for all platforms are here: http://ipyrad.readthedocs.io/installation.html" ] }, { "cell_type": "code", "execution_count": 69, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "/home/iovercast/manuscript-analysis//miniconda/bin:/home/iovercast/opt/miniconda/bin:/home/iovercast/opt/bin:/cm/shared/apps/slurm/14.11.6/sbin:/cm/shared/apps/slurm/14.11.6/bin:/cm/local/apps/gcc/5.1.0/bin:/usr/local/bin:/usr/bin:/usr/local/sbin:/usr/sbin:/sbin:/usr/sbin:/home/iovercast/.local/bin:/home/iovercast/bin\n", "PREFIX=/home/iovercast/manuscript-analysis/miniconda\n", "installing: _cache-0.0-py27_x0 ...\n", "installing: python-2.7.11-0 ...\n", "installing: conda-env-2.4.5-py27_0 ...\n", "installing: openssl-1.0.2g-0 ...\n", "installing: pycosat-0.6.1-py27_0 ...\n", "installing: pyyaml-3.11-py27_1 ...\n", "installing: readline-6.2-2 ...\n", "installing: requests-2.9.1-py27_0 ...\n", "installing: sqlite-3.9.2-0 ...\n", "installing: tk-8.5.18-0 ...\n", "installing: yaml-0.1.6-0 ...\n", "installing: zlib-1.2.8-0 ...\n", "installing: conda-4.0.5-py27_0 ...\n", "installing: pycrypto-2.6.1-py27_0 ...\n", "installing: pip-8.1.1-py27_1 ...\n", "installing: wheel-0.29.0-py27_0 ...\n", "installing: setuptools-20.3-py27_0 ...\n", "creating default environment...\n", "installation finished.\n", "Fetching package metadata: ....\n", ".Solving package specifications: .........\n", "\n", "Package plan for installation in environment /home/iovercast/manuscript-analysis/miniconda:\n", "\n", "The following packages will be downloaded:\n", "\n", " package | build\n", " ---------------------------|-----------------\n", " sqlite-3.13.0 | 0 4.0 MB\n", " python-2.7.12 | 1 12.1 MB\n", " conda-env-2.5.1 | py27_0 26 KB\n", " ruamel_yaml-0.11.14 | py27_0 352 KB\n", " conda-4.1.11 | py27_0 201 KB\n", " ------------------------------------------------------------\n", " Total: 16.6 MB\n", "\n", "The following NEW packages will be INSTALLED:\n", "\n", " ruamel_yaml: 0.11.14-py27_0\n", "\n", "The following packages will be UPDATED:\n", "\n", " conda: 4.0.5-py27_0 --> 4.1.11-py27_0 \n", " conda-env: 2.4.5-py27_0 --> 2.5.1-py27_0 \n", " python: 2.7.11-0 --> 2.7.12-1 \n", " sqlite: 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metadata .........\n", "Solving package specifications: ..........\n", "\n", "Package plan for installation in environment /home/iovercast/manuscript-analysis/miniconda:\n", "\n", "The following packages will be downloaded:\n", "\n", " package | build\n", " ---------------------------|-----------------\n", " libffi-3.2.1 | 0 36 KB\n", " libgfortran-3.0.0 | 1 281 KB\n", " libsodium-1.0.10 | 0 1.2 MB\n", " mkl-11.3.3 | 0 122.1 MB\n", " mpich2-1.4.1p1 | 0 2.6 MB\n", " pixman-0.32.6 | 0 2.4 MB\n", " glib-2.43.0 | 1 5.4 MB\n", " hdf5-1.8.17 | 1 1.9 MB\n", " libpng-1.6.22 | 0 214 KB\n", " libxml2-2.9.2 | 0 4.2 MB\n", " zeromq-4.1.4 | 0 4.1 MB\n", " alabaster-0.7.8 | py27_0 18 KB\n", " backports-1.0 | py27_0 1 KB\n", " backports_abc-0.4 | py27_0 5 KB\n", " conda-env-2.5.2 | py27_0 26 KB\n", " cython-0.24.1 | py27_0 5.3 MB\n", " decorator-4.0.10 | py27_0 12 KB\n", " docutils-0.12 | py27_2 637 KB\n", " enum34-1.1.6 | py27_0 53 KB\n", " freetype-2.5.5 | 1 2.5 MB\n", " funcsigs-1.0.2 | py27_0 18 KB\n", " functools32-3.2.3.2 | py27_0 15 KB\n", " futures-3.0.5 | py27_0 18 KB\n", " imagesize-0.7.1 | py27_0 3 KB\n", " ipython_genutils-0.1.0 | py27_0 32 KB\n", " markupsafe-0.23 | py27_2 31 KB\n", " mistune-0.7.2 | py27_0 570 KB\n", " mpi4py-2.0.0 | py27_1 1.7 MB\n", " numpy-1.11.1 | py27_0 6.2 MB\n", " path.py-8.2.1 | py27_0 45 KB\n", " ptyprocess-0.5.1 | py27_0 19 KB\n", " pygments-2.1.3 | py27_0 1.2 MB\n", " pytz-2016.6.1 | py27_0 178 KB\n", " pyzmq-15.4.0 | py27_0 705 KB\n", " simplegeneric-0.8.1 | py27_1 7 KB\n", " sip-4.18 | py27_0 264 KB\n", " six-1.10.0 | py27_0 16 KB\n", " snowballstemmer-1.2.1 | py27_0 73 KB\n", " sphinx_rtd_theme-0.1.9 | py27_0 628 KB\n", " wcwidth-0.1.7 | py27_0 21 KB\n", " babel-2.3.3 | py27_0 4.9 MB\n", " clyent-1.2.2 | py27_0 15 KB\n", " configparser-3.5.0 | py27_0 31 KB\n", " fontconfig-2.11.1 | 6 405 KB\n", " get_terminal_size-1.0.0 | py27_0 4 KB\n", " h5py-2.6.0 | np111py27_2 2.4 MB\n", " jinja2-2.8 | py27_1 264 KB\n", " jsonschema-2.5.1 | py27_0 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668 KB\n", " ipyparallel-5.2.0 | py27_0 267 KB\n", " jupyter_console-5.0.0 | py27_0 28 KB\n", " notebook-4.2.2 | py27_0 5.0 MB\n", " qt-4.8.7 | 4 32.7 MB\n", " pyqt-4.11.4 | py27_4 3.3 MB\n", " widgetsnbextension-1.2.6 | py27_0 1.1 MB\n", " qtconsole-4.2.1 | py27_0 159 KB\n", " ipyrad-0.3.36 | 4 16.6 MB ipyrad\n", " ipywidgets-5.2.2 | py27_0 58 KB\n", " jupyter-1.0.0 | py27_3 3 KB\n", " nb_anacondacloud-1.2.0 | py27_0 23 KB\n", " nb_conda_kernels-2.0.0 | py27_0 30 KB\n", " nb_conda-2.0.0 | py27_0 28 KB\n", " _nb_ext_conf-0.3.0 | py27_0 956 B\n", " nbpresent-3.0.2 | py27_0 463 KB\n", " ------------------------------------------------------------\n", " Total: 292.5 MB\n", "\n", "The following NEW packages will be INSTALLED:\n", "\n", " _nb_ext_conf: 0.3.0-py27_0 \n", " alabaster: 0.7.8-py27_0 \n", " anaconda-client: 1.5.1-py27_0 \n", " babel: 2.3.3-py27_0 \n", " backports: 1.0-py27_0 \n", " backports_abc: 0.4-py27_0 \n", " cairo: 1.12.18-6 \n", " clyent: 1.2.2-py27_0 \n", " configparser: 3.5.0-py27_0 \n", " cython: 0.24.1-py27_0 \n", " decorator: 4.0.10-py27_0 \n", " docutils: 0.12-py27_2 \n", " entrypoints: 0.2.2-py27_0 \n", " enum34: 1.1.6-py27_0 \n", " fontconfig: 2.11.1-6 \n", " freetype: 2.5.5-1 \n", " funcsigs: 1.0.2-py27_0 \n", " functools32: 3.2.3.2-py27_0 \n", " futures: 3.0.5-py27_0 \n", " get_terminal_size: 1.0.0-py27_0 \n", " glib: 2.43.0-1 \n", " h5py: 2.6.0-np111py27_2 \n", " harfbuzz: 0.9.39-1 \n", " hdf5: 1.8.17-1 \n", " imagesize: 0.7.1-py27_0 \n", " ipykernel: 4.4.1-py27_0 \n", " ipyparallel: 5.2.0-py27_0 \n", " ipyrad: 0.3.36-4 ipyrad\n", " ipython: 5.1.0-py27_0 \n", " ipython_genutils: 0.1.0-py27_0 \n", " ipywidgets: 5.2.2-py27_0 \n", " jinja2: 2.8-py27_1 \n", " jsonschema: 2.5.1-py27_0 \n", " jupyter: 1.0.0-py27_3 \n", " jupyter_client: 4.3.0-py27_0 \n", " jupyter_console: 5.0.0-py27_0 \n", " jupyter_core: 4.1.1-py27_0 \n", " libffi: 3.2.1-0 \n", " libgfortran: 3.0.0-1 \n", " libpng: 1.6.22-0 \n", " libsodium: 1.0.10-0 \n", " libxml2: 2.9.2-0 \n", " llvmlite: 0.13.0-py27_0 \n", " markupsafe: 0.23-py27_2 \n", " mistune: 0.7.2-py27_0 \n", " mkl: 11.3.3-0 \n", " mpi4py: 2.0.0-py27_1 \n", " mpich2: 1.4.1p1-0 \n", " nb_anacondacloud: 1.2.0-py27_0 \n", " nb_conda: 2.0.0-py27_0 \n", " nb_conda_kernels: 2.0.0-py27_0 \n", " nbconvert: 4.2.0-py27_0 \n", " nbformat: 4.1.0-py27_0 \n", " nbpresent: 3.0.2-py27_0 \n", " notebook: 4.2.2-py27_0 \n", " numba: 0.28.1-np111py27_0 \n", " numpy: 1.11.1-py27_0 \n", " pandas: 0.18.1-np111py27_0 \n", " pango: 1.39.0-1 \n", " path.py: 8.2.1-py27_0 \n", " pathlib2: 2.1.0-py27_0 \n", " pexpect: 4.0.1-py27_0 \n", " pickleshare: 0.7.3-py27_0 \n", " pixman: 0.32.6-0 \n", " prompt_toolkit: 1.0.3-py27_0 \n", " ptyprocess: 0.5.1-py27_0 \n", " pygments: 2.1.3-py27_0 \n", " pyqt: 4.11.4-py27_4 \n", " python-dateutil: 2.5.3-py27_0 \n", " pytz: 2016.6.1-py27_0 \n", " pyzmq: 15.4.0-py27_0 \n", " qt: 4.8.7-4 \n", " qtconsole: 4.2.1-py27_0 \n", " scipy: 0.18.0-np111py27_0 \n", " simplegeneric: 0.8.1-py27_1 \n", " singledispatch: 3.4.0.3-py27_0 \n", " sip: 4.18-py27_0 \n", " six: 1.10.0-py27_0 \n", " snowballstemmer: 1.2.1-py27_0 \n", " sphinx: 1.4.1-py27_0 \n", " sphinx_rtd_theme: 0.1.9-py27_0 \n", " ssl_match_hostname: 3.4.0.2-py27_1 \n", " terminado: 0.6-py27_0 \n", " tornado: 4.4.1-py27_0 \n", " traitlets: 4.2.2-py27_0 \n", " wcwidth: 0.1.7-py27_0 \n", " widgetsnbextension: 1.2.6-py27_0 \n", " zeromq: 4.1.4-0 \n", "\n", "The following packages will be UPDATED:\n", "\n", " conda-env: 2.5.1-py27_0 --> 2.5.2-py27_0\n", "\n", "Fetching packages ...\n", "libffi-3.2.1-0 0% | | ETA: --:--:-- 0.00 B/s\r", "libffi-3.2.1-0 44% |############# | ETA: 0:00:00 1.31 MB/s\r", "libffi-3.2.1-0 88% |########################### | ETA: 0:00:00 1.57 MB/s\r", "libffi-3.2.1-0 100% |###############################| ETA: 0:00:00 1.77 MB/s\r", "libffi-3.2.1-0 100% |###############################| Time: 0:00:00 1.68 MB/s\r\n", "libgfortran-3. 0% | | ETA: --:--:-- 0.00 B/s\r", "libgfortran-3. 5% |# | Time: 0:00:00 1.43 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We do this\n", "## so the analysis pipeline is totally self contained, it doesn't read to\n", "## or write from anywwhere but the WORK_DIR.\n", "## There's probably a magic way to do this but i can't figure it out\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"WORK_DIR=$1\"\n", "\n", "## Fetch the latest miniconda installer\n", "wget --quiet https://repo.continuum.io/miniconda/Miniconda-latest-Linux-x86_64.sh\n", "\n", "## Install miniconda silently to the work directory.\n", "## -b means \"batch\" mode, -f is force overwrite and -p is the install dir\n", "bash Miniconda-latest-Linux-x86_64.sh -b -f -p $WORK_DIR/miniconda\n", "\n", "conda update -y conda ## updates conda\n", "conda install -y -c ipyrad ipyrad ## installs the latest release (silently `-y`)" ] }, { "cell_type": "code", "execution_count": 70, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "/home/iovercast/manuscript-analysis/miniconda/bin/ipyrad\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"\n", "which ipyrad" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## Prereqs - Install pyrad\n" ] }, { "cell_type": "code", "execution_count": 71, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "Fetching package metadata .......\n", "Solving package specifications: ..........\n", "\n", "# All requested packages already installed.\n", "# packages in environment at /home/iovercast/manuscript-analysis/miniconda:\n", "#\n", "numpy 1.11.1 py27_0 \n", "scipy 0.18.0 np111py27_0 \n", "muscle3.8.31_i86linux64\n", "running install\n", "running bdist_egg\n", "running egg_info\n", "creating pyrad.egg-info\n", "writing requirements to pyrad.egg-info/requires.txt\n", "writing pyrad.egg-info/PKG-INFO\n", "writing top-level names to pyrad.egg-info/top_level.txt\n", "writing dependency_links to pyrad.egg-info/dependency_links.txt\n", "writing entry points to pyrad.egg-info/entry_points.txt\n", "writing manifest file 'pyrad.egg-info/SOURCES.txt'\n", "reading manifest file 'pyrad.egg-info/SOURCES.txt'\n", "writing manifest file 'pyrad.egg-info/SOURCES.txt'\n", "installing library code to build/bdist.linux-x86_64/egg\n", "running install_lib\n", "running build_py\n", "creating build\n", "creating build/lib\n", "creating build/lib/pyrad\n", "copying pyrad/Dtest.py -> build/lib/pyrad\n", "copying pyrad/Dtest_5.py -> build/lib/pyrad\n", "copying pyrad/Dtest_foil.py -> build/lib/pyrad\n", "copying pyrad/H_err_dp.py -> build/lib/pyrad\n", "copying pyrad/__init__.py -> build/lib/pyrad\n", "copying pyrad/alignable.py -> build/lib/pyrad\n", "copying pyrad/cluster7dp.py -> build/lib/pyrad\n", "copying pyrad/cluster_cons7_shuf.py -> build/lib/pyrad\n", "copying pyrad/consens_pairs.py -> build/lib/pyrad\n", "copying pyrad/consensdp.py -> build/lib/pyrad\n", "copying pyrad/createfile.py -> build/lib/pyrad\n", "copying pyrad/editraw_merges.py -> build/lib/pyrad\n", "copying pyrad/editraw_pairs.py -> build/lib/pyrad\n", "copying pyrad/editraw_rads.py -> build/lib/pyrad\n", "copying pyrad/loci2SNP.py -> build/lib/pyrad\n", "copying pyrad/loci2gphocs.py -> build/lib/pyrad\n", "copying pyrad/loci2mig.py -> build/lib/pyrad\n", "copying pyrad/loci2phynex.py -> build/lib/pyrad\n", "copying pyrad/loci2treemix.py -> build/lib/pyrad\n", "copying pyrad/loci2vcf.py -> build/lib/pyrad\n", "copying pyrad/overlapcheck.py -> build/lib/pyrad\n", "copying pyrad/potpour.py -> build/lib/pyrad\n", "copying pyrad/pyRAD.py -> build/lib/pyrad\n", "copying pyrad/sortandcheck2.py -> build/lib/pyrad\n", "copying pyrad/tier2clust.py -> build/lib/pyrad\n", "creating build/bdist.linux-x86_64\n", "creating build/bdist.linux-x86_64/egg\n", "creating build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/Dtest.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/Dtest_5.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/Dtest_foil.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/H_err_dp.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/__init__.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/alignable.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/cluster7dp.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/cluster_cons7_shuf.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/consens_pairs.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/consensdp.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/createfile.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/editraw_merges.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/editraw_pairs.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/editraw_rads.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/loci2SNP.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/loci2gphocs.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/loci2mig.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/loci2phynex.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/loci2treemix.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/loci2vcf.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/overlapcheck.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/potpour.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/pyRAD.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/sortandcheck2.py -> build/bdist.linux-x86_64/egg/pyrad\n", "copying build/lib/pyrad/tier2clust.py -> build/bdist.linux-x86_64/egg/pyrad\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/Dtest.py to Dtest.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/Dtest_5.py to Dtest_5.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/Dtest_foil.py to Dtest_foil.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/H_err_dp.py to H_err_dp.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/__init__.py to __init__.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/alignable.py to alignable.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/cluster7dp.py to cluster7dp.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/cluster_cons7_shuf.py to cluster_cons7_shuf.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/consens_pairs.py to consens_pairs.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/consensdp.py to consensdp.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/createfile.py to createfile.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/editraw_merges.py to editraw_merges.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/editraw_pairs.py to editraw_pairs.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/editraw_rads.py to editraw_rads.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/loci2SNP.py to loci2SNP.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/loci2gphocs.py to loci2gphocs.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/loci2mig.py to loci2mig.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/loci2phynex.py to loci2phynex.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/loci2treemix.py to loci2treemix.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/loci2vcf.py to loci2vcf.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/overlapcheck.py to overlapcheck.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/potpour.py to potpour.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/pyRAD.py to pyRAD.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/sortandcheck2.py to sortandcheck2.pyc\n", "byte-compiling build/bdist.linux-x86_64/egg/pyrad/tier2clust.py to tier2clust.pyc\n", "creating build/bdist.linux-x86_64/egg/EGG-INFO\n", "copying pyrad.egg-info/PKG-INFO -> build/bdist.linux-x86_64/egg/EGG-INFO\n", "copying pyrad.egg-info/SOURCES.txt -> build/bdist.linux-x86_64/egg/EGG-INFO\n", "copying pyrad.egg-info/dependency_links.txt -> build/bdist.linux-x86_64/egg/EGG-INFO\n", "copying pyrad.egg-info/entry_points.txt -> build/bdist.linux-x86_64/egg/EGG-INFO\n", "copying pyrad.egg-info/requires.txt -> build/bdist.linux-x86_64/egg/EGG-INFO\n", "copying pyrad.egg-info/top_level.txt -> build/bdist.linux-x86_64/egg/EGG-INFO\n", "creating dist\n", "creating 'dist/pyrad-3.0.66-py2.7.egg' and adding 'build/bdist.linux-x86_64/egg' to it\n", "removing 'build/bdist.linux-x86_64/egg' (and everything under it)\n", "Processing pyrad-3.0.66-py2.7.egg\n", "Copying pyrad-3.0.66-py2.7.egg to /home/iovercast/manuscript-analysis/miniconda/lib/python2.7/site-packages\n", "Adding pyrad 3.0.66 to easy-install.pth file\n", "Installing pyrad script to /home/iovercast/manuscript-analysis/miniconda/bin\n", "\n", "Installed /home/iovercast/manuscript-analysis/miniconda/lib/python2.7/site-packages/pyrad-3.0.66-py2.7.egg\n", "Processing dependencies for pyrad==3.0.66\n", "Searching for scipy==0.18.0\n", "Best match: scipy 0.18.0\n", "Adding scipy 0.18.0 to easy-install.pth file\n", "\n", "Using /home/iovercast/manuscript-analysis/miniconda/lib/python2.7/site-packages\n", "Searching for numpy==1.11.1\n", "Best match: numpy 1.11.1\n", "Adding numpy 1.11.1 to easy-install.pth file\n", "\n", "Using /home/iovercast/manuscript-analysis/miniconda/lib/python2.7/site-packages\n", "Finished processing dependencies for pyrad==3.0.66\n", "/home/iovercast/manuscript-analysis/miniconda/bin/muscle\n", "/home/iovercast/manuscript-analysis/miniconda/bin/vsearch\n", "/home/iovercast/manuscript-analysis/miniconda/bin/pyrad\n" ] }, { "name": "stderr", "output_type": "stream", "text": [ "Using Anaconda API: https://api.anaconda.org\n", "--2016-09-01 17:05:22-- http://www.drive5.com/muscle/downloads3.8.31/muscle3.8.31_i86linux64.tar.gz\n", "Resolving www.drive5.com (www.drive5.com)... 199.195.116.69\n", "Connecting to www.drive5.com (www.drive5.com)|199.195.116.69|:80... connected.\n", "HTTP request sent, awaiting response... 200 OK\n", "Length: 487906 (476K) [application/x-gzip]\n", "Saving to: ‘muscle3.8.31_i86linux64.tar.gz.1’\n", "\n", " 0K .......... .......... .......... .......... .......... 10% 961K 0s\n", " 50K .......... .......... .......... .......... .......... 20% 1.93M 0s\n", " 100K .......... .......... .......... .......... .......... 31% 1.93M 0s\n", " 150K .......... .......... .......... .......... .......... 41% 2.72M 0s\n", " 200K .......... .......... .......... .......... .......... 52% 3.05M 0s\n", " 250K .......... .......... .......... .......... .......... 62% 2.52M 0s\n", " 300K .......... .......... .......... .......... .......... 73% 2.54M 0s\n", " 350K .......... .......... .......... .......... .......... 83% 2.50M 0s\n", " 400K .......... .......... .......... .......... .......... 94% 2.72M 0s\n", " 450K .......... .......... ...... 100% 136M=0.2s\n", "\n", "2016-09-01 17:05:23 (2.19 MB/s) - ‘muscle3.8.31_i86linux64.tar.gz.1’ saved [487906/487906]\n", "\n", "Invalid command line option \"h\"\n", "\n", "MUSCLE v3.8.31 by Robert C. Edgar\n", "\n", "http://www.drive5.com/muscle\n", "This software is donated to the public domain.\n", "Please cite: Edgar, R.C. Nucleic Acids Res 32(5), 1792-97.\n", "\n", "\n", "Basic usage\n", "\n", " muscle -in -out \n", "\n", "Common options (for a complete list please see the User Guide):\n", "\n", " -in Input file in FASTA format (default stdin)\n", " -out Output alignment in FASTA format (default stdout)\n", " -diags Find diagonals (faster for similar sequences)\n", " -maxiters Maximum number of iterations (integer, default 16)\n", " -maxhours Maximum time to iterate in hours (default no limit)\n", " -html Write output in HTML format (default FASTA)\n", " -msf Write output in GCG MSF format (default FASTA)\n", " -clw Write output in CLUSTALW format (default FASTA)\n", " -clwstrict As -clw, with 'CLUSTAL W (1.81)' header\n", " -log[a] Log to file (append if -loga, overwrite if -log)\n", " -quiet Do not write progress messages to stderr\n", " -version Display version information and exit\n", "\n", "Without refinement (very fast, avg accuracy similar to T-Coffee): -maxiters 2\n", "Fastest possible (amino acids): -maxiters 1 -diags -sv -distance1 kbit20_3\n", "Fastest possible (nucleotides): -maxiters 1 -diags\n", "--2016-09-01 17:05:23-- https://github.com/torognes/vsearch/releases/download/v2.0.3/vsearch-2.0.3-linux-x86_64.tar.gz\n", "Resolving github.com (github.com)... 192.30.253.112\n", "Connecting to github.com (github.com)|192.30.253.112|:443... connected.\n", "HTTP request sent, awaiting response... 302 Found\n", "Location: https://github-cloud.s3.amazonaws.com/releases/19848353/7c7b511c-58d2-11e6-80a9-666723312d7e.gz?X-Amz-Algorithm=AWS4-HMAC-SHA256&X-Amz-Credential=AKIAISTNZFOVBIJMK3TQ%2F20160901%2Fus-east-1%2Fs3%2Faws4_request&X-Amz-Date=20160901T210523Z&X-Amz-Expires=300&X-Amz-Signature=513347468ae6fa6e2dc02bc4b4ff4df767b9a16fdb731254d96817663b549efb&X-Amz-SignedHeaders=host&actor_id=0&response-content-disposition=attachment%3B%20filename%3Dvsearch-2.0.3-linux-x86_64.tar.gz&response-content-type=application%2Foctet-stream [following]\n", "--2016-09-01 17:05:23-- https://github-cloud.s3.amazonaws.com/releases/19848353/7c7b511c-58d2-11e6-80a9-666723312d7e.gz?X-Amz-Algorithm=AWS4-HMAC-SHA256&X-Amz-Credential=AKIAISTNZFOVBIJMK3TQ%2F20160901%2Fus-east-1%2Fs3%2Faws4_request&X-Amz-Date=20160901T210523Z&X-Amz-Expires=300&X-Amz-Signature=513347468ae6fa6e2dc02bc4b4ff4df767b9a16fdb731254d96817663b549efb&X-Amz-SignedHeaders=host&actor_id=0&response-content-disposition=attachment%3B%20filename%3Dvsearch-2.0.3-linux-x86_64.tar.gz&response-content-type=application%2Foctet-stream\n", "Resolving github-cloud.s3.amazonaws.com (github-cloud.s3.amazonaws.com)... 54.231.40.51\n", "Connecting to github-cloud.s3.amazonaws.com (github-cloud.s3.amazonaws.com)|54.231.40.51|:443... connected.\n", "HTTP request sent, awaiting response... 200 OK\n", "Length: 487583 (476K) [application/octet-stream]\n", "Saving to: ‘vsearch-2.0.3-linux-x86_64.tar.gz’\n", "\n", " 0K .......... .......... .......... .......... .......... 10% 1.04M 0s\n", " 50K .......... .......... .......... .......... .......... 21% 1.33M 0s\n", " 100K .......... .......... .......... .......... .......... 31% 1.34M 0s\n", " 150K .......... .......... .......... .......... .......... 42% 1.32M 0s\n", " 200K .......... .......... .......... .......... .......... 52% 1.79M 0s\n", " 250K .......... .......... .......... .......... .......... 63% 1.40M 0s\n", " 300K .......... .......... .......... .......... .......... 73% 1.76M 0s\n", " 350K .......... .......... .......... .......... .......... 84% 2.41M 0s\n", " 400K .......... .......... .......... .......... .......... 94% 1.86M 0s\n", " 450K .......... .......... ...... 100% 2.86M=0.3s\n", "\n", "2016-09-01 17:05:24 (1.54 MB/s) - ‘vsearch-2.0.3-linux-x86_64.tar.gz’ saved [487583/487583]\n", "\n", "Cloning into 'pyrad'...\n", "zip_safe flag not set; analyzing archive contents...\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"WORK_DIR=$1\"\n", "\n", "## Fetch and install prerequisites. This takes a few minutes so be patient\n", "\n", "## Should be unnecessary because numpy and scipy already installed by conda\n", "conda install numpy scipy\n", "wget http://www.drive5.com/muscle/downloads3.8.31/muscle3.8.31_i86linux64.tar.gz\n", "tar -xvzf muscle*.tar.gz\n", "\n", "## This unpacks the muscle binary in the current working directory\n", "## You can test it to see if it runs:\n", "./muscle3.8.31_i86linux64 -h\n", "\n", "## Copy to miniconda/bin so it will be in your path\n", "## Maybe not technically 'correct' but it'll work for our purposes\n", "cp muscle3.8.31_i86linux64 $WORK_DIR/miniconda/bin/muscle\n", "\n", "## Download and install vsearch\n", "wget https://github.com/torognes/vsearch/releases/download/v2.0.3/vsearch-2.0.3-linux-x86_64.tar.gz\n", "tar xzf vsearch-2.0.3-linux-x86_64.tar.gz\n", "cp vsearch-2.0.3-linux-x86_64/bin/vsearch $WORK_DIR/miniconda/bin/vsearch\n", "\n", "## Fetch pyrad source from the git repository\n", "git clone https://github.com/dereneaton/pyrad.git\n", "cd pyrad\n", "\n", "## If you are using anaconda you can simply run setup.py to\n", "## install pyrad into your conda environment\n", "python setup.py install\n", "\n", "which muscle\n", "which vsearch\n", "which pyrad" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "### Prereqs - Download and install stacks\n", "\n", "**NB:** Stacks is trickier to build on OSX, I had to compile it on a different \n", "machine and then scp the binaries to the box I was working on.\n", "\n", "Stacks is picky about where stuff installs to. If you don't have permission to install to /usr/local (most HPC systems) then you need to provide the `--prefix` argument to `./configure`" ] }, { "cell_type": "code", "execution_count": 74, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "stacks-1.42/\n", "stacks-1.42/acinclude.m4\n", "stacks-1.42/aclocal.m4\n", "stacks-1.42/autogen.sh\n", "stacks-1.42/ChangeLog\n", "stacks-1.42/config/\n", "stacks-1.42/config.h.in\n", "stacks-1.42/configure\n", "stacks-1.42/configure.ac\n", "stacks-1.42/htslib/\n", "stacks-1.42/INSTALL\n", "stacks-1.42/LICENSE\n", "stacks-1.42/Makefile.am\n", "stacks-1.42/Makefile.in\n", "stacks-1.42/php/\n", "stacks-1.42/README\n", "stacks-1.42/scripts/\n", "stacks-1.42/sql/\n", "stacks-1.42/src/\n", "stacks-1.42/tests/\n", "stacks-1.42/tests/kmer_filter.t\n", "stacks-1.42/tests/process_radtags.t\n", "stacks-1.42/tests/pstacks.t\n", "stacks-1.42/tests/ustacks.t\n", "stacks-1.42/src/aln_utils.cc\n", "stacks-1.42/src/aln_utils.h\n", "stacks-1.42/src/BamI.h\n", "stacks-1.42/src/BamUnalignedI.h\n", "stacks-1.42/src/bootstrap.h\n", "stacks-1.42/src/BowtieI.h\n", "stacks-1.42/src/BustardI.h\n", "stacks-1.42/src/catalog_utils.cc\n", "stacks-1.42/src/catalog_utils.h\n", "stacks-1.42/src/clean.cc\n", "stacks-1.42/src/clean.h\n", "stacks-1.42/src/clone_filter.cc\n", "stacks-1.42/src/clone_filter.h\n", "stacks-1.42/src/cmb.cc\n", "stacks-1.42/src/cmb.h\n", "stacks-1.42/src/constants.h\n", "stacks-1.42/src/cstacks.cc\n", "stacks-1.42/src/cstacks.h\n", "stacks-1.42/src/DNANSeq.cc\n", "stacks-1.42/src/DNANSeq.h\n", "stacks-1.42/src/DNASeq.cc\n", "stacks-1.42/src/DNASeq.h\n", "stacks-1.42/src/estacks.cc\n", "stacks-1.42/src/estacks.h\n", "stacks-1.42/src/export_formats.cc\n", "stacks-1.42/src/export_formats.h\n", "stacks-1.42/src/FastaI.h\n", "stacks-1.42/src/FastqI.h\n", "stacks-1.42/src/file_io.cc\n", "stacks-1.42/src/file_io.h\n", "stacks-1.42/src/GappedAln.h\n", 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"stacks-1.42/htslib/cram/._sam_header.c\n", "stacks-1.42/htslib/cram/sam_header.c\n", "stacks-1.42/htslib/cram/._sam_header.h\n", "stacks-1.42/htslib/cram/sam_header.h\n", "stacks-1.42/htslib/cram/._string_alloc.c\n", "stacks-1.42/htslib/cram/string_alloc.c\n", "stacks-1.42/htslib/cram/._string_alloc.h\n", "stacks-1.42/htslib/cram/string_alloc.h\n", "stacks-1.42/htslib/cram/._thread_pool.c\n", "stacks-1.42/htslib/cram/thread_pool.c\n", "stacks-1.42/htslib/cram/._thread_pool.h\n", "stacks-1.42/htslib/cram/thread_pool.h\n", "stacks-1.42/htslib/cram/._vlen.c\n", "stacks-1.42/htslib/cram/vlen.c\n", "stacks-1.42/htslib/cram/._vlen.h\n", "stacks-1.42/htslib/cram/vlen.h\n", "stacks-1.42/htslib/cram/._zfio.c\n", "stacks-1.42/htslib/cram/zfio.c\n", "stacks-1.42/htslib/cram/._zfio.h\n", "stacks-1.42/htslib/cram/zfio.h\n", "stacks-1.42/config/compile\n", "stacks-1.42/config/depcomp\n", "stacks-1.42/config/install-sh\n", "stacks-1.42/config/missing\n", "stacks-1.42/config/test-driver\n", "checking 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GNU\n", "checking dependency style of g++... gcc3\n", "checking for gcc... gcc\n", "checking whether we are using the GNU C compiler... yes\n", "checking whether gcc accepts -g... yes\n", "checking for gcc option to accept ISO C89... none needed\n", "checking whether gcc understands -c and -o together... yes\n", "checking dependency style of gcc... gcc3\n", "checking whether g++ supports C++11 features by default... no\n", "checking whether g++ supports C++11 features with -std=gnu++11... yes\n", "checking for omp_set_num_threads in -lgomp... yes\n", "checking for gzread in -lz... yes\n", "checking for dirent.h that defines DIR... yes\n", "checking for library containing opendir... none required\n", "checking how to run the C preprocessor... gcc -E\n", "checking for grep that handles long lines and -e... /usr/bin/grep\n", "checking for egrep... /usr/bin/grep -E\n", "checking for ANSI C header files... yes\n", "checking for sys/types.h... yes\n", "checking for sys/stat.h... yes\n", "checking for stdlib.h... yes\n", "checking for string.h... yes\n", "checking for memory.h... yes\n", "checking for strings.h... yes\n", "checking for inttypes.h... yes\n", "checking for stdint.h... yes\n", "checking for unistd.h... yes\n", "checking float.h usability... yes\n", "checking float.h presence... yes\n", "checking for float.h... yes\n", "checking limits.h usability... yes\n", "checking limits.h presence... yes\n", "checking for limits.h... yes\n", "checking for stdlib.h... 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(cached) gawk\n", "checking that generated files are newer than configure... done\n", "configure: creating ./config.status\n", "config.status: creating Makefile\n", "config.status: creating config.h\n", "config.status: executing depfiles commands\n", "make all-recursive\n", "make[1]: Entering directory `/home/iovercast/manuscript-analysis/stacks-1.42'\n", "Making all in htslib\n", "make[2]: Entering directory `/home/iovercast/manuscript-analysis/stacks-1.42/htslib'\n", "echo '/* Empty config.h generated by Makefile */' > config.h\n", "gcc -g -Wall -O2 -I. -c -o kfunc.o kfunc.c\n", "gcc -g -Wall -O2 -I. -c -o knetfile.o knetfile.c\n", "gcc -g -Wall -O2 -I. -c -o kstring.o kstring.c\n", "gcc -g -Wall -O2 -I. -c -o bgzf.o bgzf.c\n", "gcc -g -Wall -O2 -I. -c -o faidx.o faidx.c\n", "gcc -g -Wall -O2 -I. -c -o hfile.o hfile.c\n", "gcc -g -Wall -O2 -I. -c -o hfile_net.o hfile_net.c\n", "echo '#define HTS_VERSION \"1.3.1\"' > version.h\n", "gcc -g -Wall -O2 -I. -c -o hts.o hts.c\n", "gcc -g 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src/.deps/phasedstacks-locus.Po\n", "g++ -DHAVE_CONFIG_H -I. -fopenmp -g -O2 -std=gnu++11 -MT src/phasedstacks-input.o -MD -MP -MF src/.deps/phasedstacks-input.Tpo -c -o src/phasedstacks-input.o `test -f 'src/input.cc' || echo './'`src/input.cc\n", "mv -f src/.deps/phasedstacks-input.Tpo src/.deps/phasedstacks-input.Po\n", "g++ -DHAVE_CONFIG_H -I. -fopenmp -g -O2 -std=gnu++11 -MT src/phasedstacks-sql_utilities.o -MD -MP -MF src/.deps/phasedstacks-sql_utilities.Tpo -c -o src/phasedstacks-sql_utilities.o `test -f 'src/sql_utilities.cc' || echo './'`src/sql_utilities.cc\n", "mv -f src/.deps/phasedstacks-sql_utilities.Tpo src/.deps/phasedstacks-sql_utilities.Po\n", "g++ -DHAVE_CONFIG_H -I. -fopenmp -g -O2 -std=gnu++11 -MT src/phasedstacks-log_utils.o -MD -MP -MF src/.deps/phasedstacks-log_utils.Tpo -c -o src/phasedstacks-log_utils.o `test -f 'src/log_utils.cc' || echo './'`src/log_utils.cc\n", "mv -f src/.deps/phasedstacks-log_utils.Tpo src/.deps/phasedstacks-log_utils.Po\n", "g++ 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Leaving directory `/home/iovercast/manuscript-analysis/stacks-1.42'\n", "Making install in htslib\n", "make[1]: Entering directory `/home/iovercast/manuscript-analysis/stacks-1.42/htslib'\n", "make[1]: Nothing to be done for `install'.\n", "make[1]: Leaving directory `/home/iovercast/manuscript-analysis/stacks-1.42/htslib'\n", "Making install in .\n", "make[1]: Entering directory `/home/iovercast/manuscript-analysis/stacks-1.42'\n", "make[2]: Entering directory `/home/iovercast/manuscript-analysis/stacks-1.42'\n", " /usr/bin/mkdir -p '/home/iovercast/manuscript-analysis//miniconda/bin/bin'\n", " /usr/bin/install -c ustacks pstacks estacks cstacks sstacks rxstacks hstacks process_radtags process_shortreads kmer_filter clone_filter genotypes populations phasedstacks '/home/iovercast/manuscript-analysis//miniconda/bin/bin'\n", " /usr/bin/mkdir -p '/home/iovercast/manuscript-analysis//miniconda/bin/bin'\n", " /usr/bin/install -c scripts/denovo_map.pl scripts/ref_map.pl scripts/export_sql.pl scripts/sort_read_pairs.pl scripts/exec_velvet.pl scripts/load_sequences.pl scripts/index_radtags.pl scripts/load_radtags.pl scripts/stacks_export_notify.pl scripts/integrate_alignments.py '/home/iovercast/manuscript-analysis//miniconda/bin/bin'\n", " /usr/bin/mkdir -p '/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks'\n", " /usr/bin/mkdir -p '/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/sql'\n", " /usr/bin/install -c -m 644 sql/mysql.cnf.dist sql/catalog_index.sql sql/stacks.sql sql/tag_index.sql sql/chr_index.sql '/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/sql'\n", " /usr/bin/mkdir -p '/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php'\n", " /usr/bin/install -c -m 644 php/CatalogClass.php php/annotate_marker.php php/constants.php.dist php/index.php php/tags.php php/tag.php php/Locus.php php/catalog.php php/correct_genotypes.php php/correct_genotype.php php/export_batch.php php/last_modified.php php/version.php php/catalog_genotypes.php php/db_functions.php php/header.php php/samples.php php/stacks_functions.php php/view_sequence.php php/sequence_blast.php php/pop_view.php php/sumstat_view.php php/hapstat_view.php php/fst_view.php php/phist_view.php php/stack_view.php php/population_view.js php/ajax.js php/annotate.js php/stacks.js php/export.js php/stacks.css '/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php'\n", " /usr/bin/mkdir -p '/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php/images'\n", " /usr/bin/install -c -m 644 php/images/caret-d.png php/images/caret-u.png php/images/excel_icon.png php/images/l-arrow-disabled.png php/images/l-arrow.png php/images/r-arrow-disabled.png php/images/r-arrow.png php/images/stacks_bg.png php/images/stacks_logo_rev_small.png '/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php/images'\n", "make install-data-hook\n", "make[3]: Entering directory `/home/iovercast/manuscript-analysis/stacks-1.42'\n", "sed -e 's,_VERSION_,1.42,' -e 's,_BINDIR_,/home/iovercast/manuscript-analysis//miniconda/bin/bin/,g' -e 's,_PKGDATADIR_,/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/,g' /home/iovercast/manuscript-analysis//miniconda/bin/bin/denovo_map.pl > /home/iovercast/manuscript-analysis//miniconda/bin/bin/denovo_map.pl.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/bin/denovo_map.pl.subst /home/iovercast/manuscript-analysis//miniconda/bin/bin/denovo_map.pl\n", "chmod +x /home/iovercast/manuscript-analysis//miniconda/bin/bin/denovo_map.pl\n", "sed -e 's,_VERSION_,1.42,' -e 's,_BINDIR_,/home/iovercast/manuscript-analysis//miniconda/bin/bin/,g' -e 's,_PKGDATADIR_,/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/,g' /home/iovercast/manuscript-analysis//miniconda/bin/bin/ref_map.pl > /home/iovercast/manuscript-analysis//miniconda/bin/bin/ref_map.pl.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/bin/ref_map.pl.subst /home/iovercast/manuscript-analysis//miniconda/bin/bin/ref_map.pl\n", "chmod +x /home/iovercast/manuscript-analysis//miniconda/bin/bin/ref_map.pl\n", "sed -e 's,_VERSION_,1.42,' -e 's,_PKGDATADIR_,/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/,g' /home/iovercast/manuscript-analysis//miniconda/bin/bin/export_sql.pl > /home/iovercast/manuscript-analysis//miniconda/bin/bin/export_sql.pl.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/bin/export_sql.pl.subst /home/iovercast/manuscript-analysis//miniconda/bin/bin/export_sql.pl\n", "chmod +x /home/iovercast/manuscript-analysis//miniconda/bin/bin/export_sql.pl\n", "sed -e 's,_VERSION_,1.42,' -e 's,_PKGDATADIR_,/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/,g' /home/iovercast/manuscript-analysis//miniconda/bin/bin/index_radtags.pl > /home/iovercast/manuscript-analysis//miniconda/bin/bin/index_radtags.pl.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/bin/index_radtags.pl.subst /home/iovercast/manuscript-analysis//miniconda/bin/bin/index_radtags.pl\n", "chmod +x /home/iovercast/manuscript-analysis//miniconda/bin/bin/index_radtags.pl\n", "sed -e 's,_VERSION_,1.42,' -e 's,_PKGDATADIR_,/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/,g' /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_radtags.pl > /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_radtags.pl.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_radtags.pl.subst /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_radtags.pl\n", "chmod +x /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_radtags.pl\n", "sed -e 's,_VERSION_,1.42,' /home/iovercast/manuscript-analysis//miniconda/bin/bin/sort_read_pairs.pl > /home/iovercast/manuscript-analysis//miniconda/bin/bin/sort_read_pairs.pl.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/bin/sort_read_pairs.pl.subst /home/iovercast/manuscript-analysis//miniconda/bin/bin/sort_read_pairs.pl\n", "chmod +x /home/iovercast/manuscript-analysis//miniconda/bin/bin/sort_read_pairs.pl\n", "sed -e 's,_VERSION_,1.42,' /home/iovercast/manuscript-analysis//miniconda/bin/bin/exec_velvet.pl > /home/iovercast/manuscript-analysis//miniconda/bin/bin/exec_velvet.pl.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/bin/exec_velvet.pl.subst /home/iovercast/manuscript-analysis//miniconda/bin/bin/exec_velvet.pl\n", "chmod +x /home/iovercast/manuscript-analysis//miniconda/bin/bin/exec_velvet.pl\n", "sed -e 's,_VERSION_,1.42,' -e 's,_PKGDATADIR_,/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/,g' /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_sequences.pl > /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_sequences.pl.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_sequences.pl.subst /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_sequences.pl\n", "chmod +x /home/iovercast/manuscript-analysis//miniconda/bin/bin/load_sequences.pl\n", "sed -e 's,_VERSION_,1.42,' -e 's,_PKGDATADIR_,/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/,g' -e 's,_BINDIR_,/home/iovercast/manuscript-analysis//miniconda/bin/bin/,g' \\\n", "\t/home/iovercast/manuscript-analysis//miniconda/bin/bin/stacks_export_notify.pl > /home/iovercast/manuscript-analysis//miniconda/bin/bin/stacks_export_notify.pl.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/bin/stacks_export_notify.pl.subst /home/iovercast/manuscript-analysis//miniconda/bin/bin/stacks_export_notify.pl\n", "chmod +x /home/iovercast/manuscript-analysis//miniconda/bin/bin/stacks_export_notify.pl\n", "sed -e 's,_PKGDATADIR_,/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/,g' -e 's,_BINDIR_,/home/iovercast/manuscript-analysis//miniconda/bin/bin/,g' \\\n", "\t/home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php/constants.php.dist > /home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php/constants.php.dist.subst\n", "mv /home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php/constants.php.dist.subst /home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php/constants.php.dist\n", "echo 1.42 > /home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php/version.php\n", "/bin/sh /home/iovercast/manuscript-analysis/stacks-1.42/config/install-sh -d -m 755 /home/iovercast/manuscript-analysis//miniconda/bin/share/stacks/php/export\n", "make[3]: Leaving directory `/home/iovercast/manuscript-analysis/stacks-1.42'\n", "make[2]: Leaving directory `/home/iovercast/manuscript-analysis/stacks-1.42'\n", "make[1]: Leaving directory `/home/iovercast/manuscript-analysis/stacks-1.42'\n", "/home/iovercast/opt/bin/process_radtags\n" ] }, { "name": "stderr", "output_type": "stream", "text": [ "--2016-09-01 17:11:52-- http://catchenlab.life.illinois.edu/stacks/source/stacks-1.42.tar.gz\n", "Resolving catchenlab.life.illinois.edu (catchenlab.life.illinois.edu)... 130.126.48.94\n", "Connecting to catchenlab.life.illinois.edu (catchenlab.life.illinois.edu)|130.126.48.94|:80... connected.\n", "HTTP request sent, awaiting response... 200 OK\n", "Length: 897951 (877K) [application/x-gzip]\n", "Saving to: ‘stacks-1.42.tar.gz.1’\n", "\n", " 0K .......... .......... .......... .......... .......... 5% 790K 1s\n", " 50K .......... .......... .......... .......... .......... 11% 1.00M 1s\n", " 100K .......... .......... .......... .......... .......... 17% 777K 1s\n", " 150K .......... .......... .......... .......... .......... 22% 637K 1s\n", " 200K .......... .......... .......... .......... .......... 28% 91.4M 1s\n", " 250K .......... .......... .......... .......... .......... 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‘stacks-1.42.tar.gz.1’ saved [897951/897951]\n", "\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"WORK_DIR=$1\"\n", "cd $WORK_DIR\n", "\n", "## 1.42 is the version that crashed during cstacks on the simdata. Julian fixed\n", "## the bug and pushed 1.43 which seems to work great.\n", "#wget http://catchenlab.life.illinois.edu/stacks/source/stacks-1.42.tar.gz\n", "wget http://catchenlab.life.illinois.edu/stacks/source/stacks-1.43.tar.gz\n", "tar -xvzf stacks-1.43.tar.gz\n", "cd stacks-1.43\n", "./configure --prefix=$WORK_DIR/miniconda\n", "make\n", "make install\n", "\n", "cd $WORK_DIR\n", "which process_radtags" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "### Prereqs - Download and install aftrRAD\n", "\n", "Lots of gnarly prereqs. Also, the `Genotype.pl` script prompts the user a couple times \n", "to verify whether or not you want to remove individuals with lots of missing data. I\n", "modified the script for the simulated data so it just passes so I can actually\n", "automate testing." ] }, { "cell_type": "code", "execution_count": 338, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "Process is interrupted.\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\" -s \"$AFTRRAD_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"WORK_DIR=$1\"; export \"AFTRRAD_DIR=$2\"\n", "cd $AFTRRAD_DIR\n", "\n", "## Get a copy of acana\n", "wget http://www.niehs.nih.gov/research/resources/assets/docs/acana_linux_x64tgz.tgz\n", "cp ACANA_DIR/ACANA .\n", "cp ACANA_DIR/dnaMatrix .\n", "\n", "## ... and mafft\n", "wget http://mafft.cbrc.jp/alignment/software/mafft-7.305-with-extensions-src.tgz\n", "tar -xvzf mafft-7.305-with-extensions-src.tgz\n", "cd mafft-7.305-with-extensions/core\n", "## This won't work on mac (`-i` flag acts different)\n", "sed -i 's/PREFIX = \\/usr\\/local/PREFIX = \\/$WORK_DIR\\/miniconda/' Makefile\n", "make; make install\n", "\n", "## ... and R\n", "## R turns out to be a complete nightmare to install if you don't have root\n", "## using brew makes it easier, but still not v straightforward\n", "## Brew installing R takes a while so be patient (>10 minutes)\n", "cd $AFTRRAD_DIR\n", "git clone https://github.com/Linuxbrew/brew.git\n", "./brew/bin/brew install homebrew/science/r\n", "## Copy the R binary to the miniconda bin dir so it will be available\n", "cp brew/bin/R $WORK_DIR/miniconda/bin/\n", "\n", "## Some hackish bullshit to get cpan working in the event you are _not_ root\n", "## which is most everybody\n", "wget -O- http://cpanmin.us | perl - -l ~/perl5 App::cpanminus local::lib\n", "eval `perl -I ~/perl5/lib/perl5 -Mlocal::lib`\n", "echo 'eval `perl -I $WORK_DIR/perl5/lib/perl5 -Mlocal::lib`' >> $WORK_DIR/.profile\n", "echo 'export MANPATH=$WORK_DIR/perl5/man:$MANPATH' >> $WORK_DIR/.profile\n", "source $WORK_DIR/.profile\n", "cpanm Parallel::ForkManager\n", "\n", "## Actually get aftrRAD\n", "git clone https://github.com/mikesovic/AftrRAD.git" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "### Install dDocent\n", "\n", "dDocent installs a bunch of business into its working directory so you have to be\n", "sure to update the PATH in a similar way to how we do it with miniconda, i.e.:\n", " \n", " %%bash -s \"$WORK_DIR\" -s \"$DDOCENT_DIR\"\n", " export PATH=\"$2/:$1/miniconda/bin:$PATH\";\n", "\n", "From the dDocent docs: \"Now if you are using a Mac computer, things get a little trickier.\" This part of the manu pipeline is only tested and known to work on linux.\n", "\n", "There is an install shell script and it tried to install lots of the deps, but several failed, which i had to install by hand (freebayes, bwa, samtools, seqtk, cd-hit-est). Drag. Freebayes turned out to be a nightmare of deps as well, so it took hours to figure out how to install. Ultimately I had to compile it on another machine and copy the binaries over. Also had to update my LD_LIBRARY_PATH to get vcflibs tools to work: \n", "\n", " export LD_LIBRARY_PATH=/home/iovercast/manuscript-analysis/dDocent/freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH" ] }, { "cell_type": "code", "execution_count": 435, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "/home/iovercast/manuscript-analysis/dDocent/:/home/iovercast/opt/miniconda/bin:/home/iovercast/opt/bin:/cm/shared/apps/slurm/14.11.6/sbin:/cm/shared/apps/slurm/14.11.6/bin:/cm/local/apps/gcc/5.1.0/bin:/usr/local/bin:/usr/bin:/usr/local/sbin:/usr/sbin:/sbin:/usr/sbin:/home/iovercast/.local/bin:/home/iovercast/bin\r\n", "dDocent 2.24 \r\n", "\r\n", "Contact jpuritz@gmail.com with any problems \r\n", "\r\n", " \r\n", "Checking for required software\r\n", "The dependency freebayes is not installed or is not in your $PATH.\r\n", "The dependency bwa is not installed or is not in your $PATH.\r\n", "The dependency samtools is not installed or is not in your $PATH.\r\n", "The dependency seqtk is not installed or is not in your $PATH.\r\n", "The dependency cd-hit-est is not installed or is not in your $PATH.\r\n", "/home/iovercast/manuscript-analysis/dDocent/dDocent: line 46: [: : integer expression expected\r\n", "The version of Samtools installed in your $PATH is not optimized for dDocent.\r\n", "Please install at least version 1.3.0\r\n", "\u0007\u0007" ] } ], "source": [ "#%%bash -s \"$WORK_DIR\" -s \"$DDOCENT_DIR\"\n", "#export PATH=\"$1/miniconda/bin:$PATH\"; export \"WORK_DIR=$1\"; export \"DDOCENT_DIR=$2\"\n", "os.chdir(WORK_DIR)\n", "#force = True\n", "force = \"\"\n", "if force:\n", " shutil.rmtree(DDOCENT_DIR)\n", " cmd = \"git clone https://github.com/jpuritz/dDocent.git\"\n", " !$cmd\n", "\n", " os.chdir(\"dDocent\")\n", " ## This will run for a long time and install a bunch of binaries to the dDocent dir\n", " cmd = \"sh install_dDocent_requirements \" + DDOCENT_DIR\n", " !$cmd\n", " cmd = \"chmod 777 {}/dDocent\".format(DDOCENT_DIR)\n", "\n", "## test\n", "cmd = \"export PATH={}:$PATH; dDocent\".format(DDOCENT_DIR)\n", "!$cmd" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "# Simulation Analysis\n", "\n", "We will use `simrrls` to generate some simulated RAD-seq data for testing. This is a program that was written by Deren Eaton and is available on github: github.com/dereneaton/simrrls.git. `simrrls` requires the python egglib module, which is somewhat painful to install. Short version:\n", "* Install egglib (painful)\n", " * Good directions: http://wjidea.github.io/2016/installEgglib.html\n", "* Install simrrls\n", " * git clone https://github.com/dereneaton/simrrls.git\n", " * cd simrrls\n", " * python setup.py install" ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": true }, "outputs": [], "source": [ "%%bash -s \"$WORK_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"WORK_DIR=$1\"\n", "cd $WORK_DIR\n", "\n", "## This is the long and painful version of installing egglib.\n", "## This is what i did, it's not super guaranteed to work on \n", "## different platforms, but it should be at least a guide to follow.\n", "\n", "## no cmake on my hpc compute node, so we have to install it ourselves\n", "## grab the silent installer and tell it to write into the \n", "## miniconda dir, since we're already adding that to path\n", "wget https://cmake.org/files/v3.6/cmake-3.6.1-Linux-x86_64.sh\n", "bash cmake-3.6.1-Linux-x86_64.sh --prefix=$WORK_DIR/miniconda --exclude-subdir\n", "cmake --version\n", "\n", "## Install bio++. Similar move here, we'll install to the miniconda\n", "## directory because we don't have any assurance of root on the box\n", "mkdir bpp\n", "cd bpp\n", "wget http://biopp.univ-montp2.fr/repos/sources/bpp-core-2.2.0.tar.gz\n", "wget http://biopp.univ-montp2.fr/repos/sources/bpp-seq-2.2.0.tar.gz\n", "wget http://biopp.univ-montp2.fr/repos/sources/bpp-popgen-2.2.0.tar.gz\n", "tar -xvzf bpp-core-2.2.0.tar.gz\n", "tar -xvzf bpp-seq-2.2.0.tar.gz\n", "tar -xvzf bpp-popgen-2.2.0.tar.gz\n", "cd bpp-core-2.2.0\n", "cmake -DCMAKE_INSTALL_PREFIX=$WORK_DIR/miniconda\n", "make\n", "make install\n", "cd ../bpp-seq-2.2.0\n", "cmake -DCMAKE_INSTALL_PREFIX=$WORK_DIR/miniconda\n", "make\n", "make install\n", "cd ../bpp-popgen-2.2.0\n", "cmake -DCMAKE_INSTALL_PREFIX=$WORK_DIR/miniconda\n", "make\n", "make install\n", "\n", "## install egglib-cpp and egglib python module\n", "## NB: simrrls requires egglib 2 \n", "wget http://mycor.nancy.inra.fr/egglib/releases/2.1.11/egglib-cpp-2.1.11.tar.gz\n", "wget http://mycor.nancy.inra.fr/egglib/releases/2.1.11/egglib-py-2.1.11.tar.gz\n", "tar -xvzf egglib-cpp-2.1.11.tar.gz\n", "tar -xvzf egglib-py-2.1.11.tar.gz\n", "cd egglib-cpp-2.1.11\n", "./configure --prefix=$WORK_DIR/miniconda\n", "make\n", "make install\n", "cd ../egglib-py-2.1.11/\n", "python setup.py build --prefix=$WORK_DIR/miniconda\n", "python setup.py install --prefix=$WORK_DIR/miniconda\n", "\n", "## Install simrrls\n", "git clone https://github.com/dereneaton/simrrls.git\n", "cd simrrls\n", "python setup.py install\n", "simrrls --help" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## Simulating different RAD-datasets\n", "Both pyRAD and stacks have undergone a lot of work since the original pyrad analysis. Because improvements have been made we want to test performance of all the current pipelines and be able to compare current to past performance. We'll follow the original pyRAD manuscript analysis (Eaton 2013) by simulating modest sized datasets with variable amounts of indels. We'll also simulate one much larger dataset. Also, because stacks has since included an option for handling gapped analysis we'll test both gapped and ungapped assembly.\n", "\n", "### Tuning simrrls indel parameter\n", "The `-I` parameter for simrrls has changed since the initial pyrad manuscript, so the we had to explore new values for this parameter that will approximate the number of indels we are after. I figured out a way to run simrrls and pipe the output to muscle to get a quick idea of the indel variation for different params:\n", "\n", " simrrls -n 1 -L 1 -I 1 -r1 $RANDOM 2>&1 | grep 0 -A 1 | tr '@' '>' | muscle | grep T | head -n 60\n", "\n", "If you run it like this then you can get an idea of how many indel bearing seqs are generated:\n", "\n", " for i in {1..50}; do simrrls -n 1 -L 1 -I .05 -r1 $RANDOM 2>&1 | grep 0 -A 1 | tr '@' '>' | muscle | grep T | head -n 40 >> rpt.txt; done\n", " grep \"-\" rpt.txt | wc -l\n", " \n", "From experimentation:\n", "\n", "-I value -- %loci w/ indels\n", "* 0.02 -- ~10%\n", "* 0.05 -- ~15%\n", "* 0.10 -- ~25%\n", "\n", "The simulated data will live in these directories:\n", "* SIM_NO_DIR = WORK_DIR/simulated_data/simno\n", "* SIM_LO_DIR = WORK_DIR/simulated_data/simlo\n", "* SIM_HI_DIR = WORK_DIR/simulated_data/simhi\n", "* SIM_LARGE_DIR = WORK_DIR/simulated_data/simlarge\n", "\n", "Timing:\n", "* 10K loci -- ~8MB -- ~ 2 minutes\n", "* 100K loci -- ~80MB -- ~ 20 minutes" ] }, { "cell_type": "code", "execution_count": 326, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time simrrls -o /home/iovercast/manuscript-analysis/simulated_data/simno/simno -ds 2 -L 10000 -I 0 -r1 1\n", "\n", "\n", "real\t1m56.606s\n", "user\t1m56.326s\n", "sys\t0m0.257s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time simrrls -o /home/iovercast/manuscript-analysis/simulated_data/simlo/simlo -ds 2 -L 10000 -I 0.02 -r1 1\n", "\n", "\n", "real\t2m5.706s\n", "user\t2m5.544s\n", "sys\t0m0.183s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time simrrls -o /home/iovercast/manuscript-analysis/simulated_data/simhi/simhi -ds 2 -L 10000 -I 0.05 -r1 1\n", "\n", "\n", "real\t2m8.086s\n", "user\t2m7.912s\n", "sys\t0m0.191s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time simrrls -o /home/iovercast/manuscript-analysis/simulated_data/simlarge/simlarge -ds 2 -L 100000 -mc 1 -r1 1\n", "\n", "real\t2m8.086s\n", "user\t2m7.912s\n", "sys\t0m0.191s\n", "\n" ] } ], "source": [ "import subprocess\n", "import shutil\n", "\n", "force = True\n", "\n", "## Directories for the simulation data\n", "SIM_NO_DIR=os.path.join(SIMULATION_DATA_DIR, \"simno\")\n", "SIM_LO_DIR=os.path.join(SIMULATION_DATA_DIR, \"simlo\")\n", "SIM_HI_DIR=os.path.join(SIMULATION_DATA_DIR, \"simhi\")\n", "SIM_LARGE_DIR=os.path.join(SIMULATION_DATA_DIR, \"simlarge\")\n", "\n", "for dir in [SIMULATION_DATA_DIR, SIM_NO_DIR, SIM_LO_DIR,\\\n", " SIM_HI_DIR, SIM_LARGE_DIR]:\n", " if force and os.path.exists(dir):\n", " shutil.rmtree(dir)\n", " if not os.path.exists(dir):\n", " os.makedirs(dir)\n", "\n", "\n", "## Tree input doesn't work right. Egglib errors.\n", "## These are the trees from pyrad v3 manu, not sure if they ever worked there either\n", "# newick = \"(((((A:2,B:2):2,C:4):4,D:8):4,(((E:2,F:2):2,G:4):4,H:8):4):4,(((I:2,J:2):2,K:4):4,L:8):8,X:16):16;\"\n", "newick = \"(((((A,B),C),D),(((E,F),G),H)),(((I,J),K),L));\"\n", "treefile = os.path.join(SIMULATION_DATA_DIR, \"simtree.txt\")\n", "with open(treefile, 'w') as outfile:\n", " outfile.write(newick)\n", "\n", "cmd = \"\"\"\\\n", "export PATH={workdir}/miniconda/bin:$PATH; time simrrls -o {odir}/{oname} -ds 2 -L 10000 -I {indels} -r1 1\n", "\"\"\"\n", "\n", "## simulate no indels\n", "call = cmd.format(workdir=WORK_DIR, odir=SIMULATION_DATA_DIR, oname=\"simno/simno\", indels=0, tree=treefile)\n", "print(call)\n", "output = subprocess.check_output(call, shell=True, stderr=subprocess.STDOUT)\n", "print(output)\n", "\n", "## simulate low indels\n", "call = cmd.format(workdir=WORK_DIR, odir=SIMULATION_DATA_DIR, oname=\"simlo/simlo\", indels=0.02)\n", "print(call)\n", "output = subprocess.check_output(call, shell=True, stderr=subprocess.STDOUT)\n", "print(output)\n", "\n", "## simulate high indels\n", "call = cmd.format(workdir=WORK_DIR, odir=SIMULATION_DATA_DIR, oname=\"simhi/simhi\", indels=0.05)\n", "print(call)\n", "output = subprocess.check_output(call, shell=True, stderr=subprocess.STDOUT)\n", "print(output)\n", "\n", "## simulate large dataset, no indels but w/ allele dropout from mutations to cutsites\n", "call = \"export PATH={workdir}/miniconda/bin:$PATH; time simrrls -o {odir}/{oname} -ds 2 -L 100000 -mc 1 -r1 1\"\\\n", " .format(workdir=WORK_DIR, odir=SIMULATION_DATA_DIR, oname=\"simlarge/simlarge\")\n", "print(call)\n", "#output = subprocess.check_output(call, shell=True, stderr=subprocess.STDOUT)\n", "print(output)" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## ipyrad simulated data assembly\n", "\n", "For simulated datasets with 10000 l00bp loci ipyrad runs in \n", "* v.0.3.41 ~6 minutes.\n", "* v.0.3.42 ~8 minutes.\n", "* v.0.4.0 ~4 minutes.\n", "* v.0.5.10 ~4 minutes.\n", "\n", "Large dataset (100000 100bp loci)\n", "* v.0.4.0 ~24 minutes." ] }, { "cell_type": "code", "execution_count": 2, "metadata": { "collapsed": false }, "outputs": [ { "data": { "text/plain": [ "'0.5.10'" ] }, "execution_count": 2, "metadata": {}, "output_type": "execute_result" } ], "source": [ "import ipyrad as ip\n", "reload(ip)\n", "ip.__version__" ] }, { "cell_type": "code", "execution_count": 3, "metadata": { "collapsed": false, "scrolled": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ " New Assembly: simhi\n", "ipyrad -p params-simhi.txt -s 1234567 --force\n", "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.5.10]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: simhi\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Demultiplexing fastq data to Samples\n", " [####################] 100% chunking large files | 0:00:00 \n", " [####################] 100% sorting reads | 0:02:11 \n", " [####################] 100% writing/compressing | 0:00:06 \n", "\n", " Step 2: Filtering reads \n", " [####################] 100% processing reads | 0:00:21 \n", "\n", " Step 3: Clustering/Mapping reads\n", " [####################] 100% dereplicating | 0:00:00 \n", " [####################] 100% clustering | 0:00:04 \n", " [####################] 100% building clusters | 0:00:05 \n", " [####################] 100% chunking | 0:00:00 \n", " [####################] 100% aligning | 0:02:33 \n", " [####################] 100% concatenating | 0:00:02 \n", "\n", " Step 4: Joint estimation of error rate and heterozygosity\n", " [####################] 100% inferring [H, E] | 0:01:45 \n", "\n", " Step 5: Consensus base calling \n", " Mean error [0.00075 sd=0.00001]\n", " Mean hetero [0.00192 sd=0.00005]\n", " [####################] 100% calculating depths | 0:00:01 \n", " [####################] 100% chunking clusters | 0:00:02 \n", " [####################] 100% consens calling | 0:01:19 \n", "\n", " Step 6: Clustering at 0.85 similarity across 12 samples\n", " [####################] 100% concat/shuffle input | 0:00:03 \n", " [####################] 100% clustering across | 0:00:06 \n", " [####################] 100% building clusters | 0:00:04 \n", " [####################] 100% aligning clusters | 0:00:23 \n", " [####################] 100% database indels | 0:00:06 \n", " [####################] 100% indexing clusters | 0:00:08 \n", " [####################] 100% building database | 0:00:09 \n", "\n", " Step 7: Filter and write output files for 12 Samples\n", " [####################] 100% filtering loci | 0:00:21 \n", " [####################] 100% building loci/stats | 0:00:03 \n", " [####################] 100% building vcf file | 0:00:07 \n", " [####################] 100% writing vcf file | 0:00:00 \n", " [####################] 100% building arrays | 0:00:05 \n", " [####################] 100% writing outfiles | 0:00:03 \n", " Outfiles written to: ~/manuscript-analysis/ipyrad/SIMDATA/simhi/simhi_outfiles\n", "\n", "\n", "real\t11m11.744s\n", "user\t1m4.042s\n", "sys\t0m2.858s\n" ] } ], "source": [ "import ipyrad as ip\n", "\n", "## Set up directory structures. change the force flag if you want to\n", "## blow everything away and restart\n", "#force = True\n", "force = \"\"\n", "IPYRAD_SIMOUT=os.path.join(IPYRAD_DIR, \"SIMDATA/\")\n", "if force and os.path.exists(IPYRAD_SIMOUT):\n", " shutil.rmtree(IPYRAD_SIMOUT)\n", "if not os.path.exists(IPYRAD_SIMOUT):\n", " os.makedirs(IPYRAD_SIMOUT)\n", "for outdir in [\"no/\", \"lo/\", \"hi/\", \"large/\"]:\n", " tmpdir = os.path.join(IPYRAD_SIMOUT, \"sim\"+outdir)\n", " if not os.path.exists(tmpdir):\n", " os.makedirs(tmpdir)\n", "\n", "## go to the ipyrad simulated data output directory\n", "#for dir in [\"no\", \"lo\", \"hi\", \"large\"]:\n", "for dir in [\"hi\"]: \n", " ## Set barcode and raw data files\n", " simout_dir = os.path.join(IPYRAD_SIMOUT, \"sim\"+dir)\n", " simdata_dir = os.path.join(SIMULATION_DATA_DIR, \"sim\"+dir)\n", " bcodes = os.path.join(simdata_dir, \"sim{}_barcodes.txt\".format(dir))\n", " fastqs = os.path.join(simdata_dir, \"sim{}_R1_.fastq.gz\".format(dir))\n", " os.chdir(simout_dir)\n", " ## Make a new assembly\n", " data = ip.Assembly(\"sim\"+dir)\n", " \n", " ## Set padata1.set_params('project_dir', \"./test_rad\")\n", " data.set_params('raw_fastq_path', fastqs)\n", " data.set_params('barcodes_path', bcodes)\n", " data.set_params('max_low_qual_bases', 4)\n", " data.set_params('filter_min_trim_len', 69)\n", " data.set_params('max_Ns_consens', (99,99))\n", " data.set_params('max_Hs_consens', (99,99))\n", " data.set_params('max_SNPs_locus', (100, 100))\n", " data.set_params('min_samples_locus', 2)\n", " data.set_params('max_Indels_locus', (99,99))\n", " data.set_params('max_shared_Hs_locus', 99)\n", " data.set_params('trim_overhang', (2,2,2,2))\n", "\n", " data.write_params(force=True)\n", "\n", " cmd = \"ipyrad -p params-sim{}.txt -s 1234567 --force\".format(dir)\n", " print(cmd)\n", " !time $cmd" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## pyrad simulated data assembly\n", "Simulated assembly approximate runtimes:\n", "\n", "* 10000 l00bp loci pyrad runs in ~11 minutes.\n", "* 100000 100bp loci pyrad runs in ~90 minutes." ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "### Set the default pyrad params file." ] }, { "cell_type": "code", "execution_count": 8, "metadata": { "collapsed": false, "scrolled": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "==** parameter inputs for pyRAD version 3.0.63 **======================== affected step ==\n", " ## 1. Working directory (all)\n", " ## 2. Loc. of non-demultiplexed files (if not line 18) (s1)\n", " ## 3. Loc. of barcode file (if not line 18) (s1)\n", "vsearch ## 4. command (or path) to call vsearch (or usearch) (s3,s6)\n", "muscle ## 5. command (or path) to call muscle (s3,s7)\n", "TGCAG ## 6. Restriction overhang (e.g., C|TGCAG -> TGCAG) (s1,s2)\n", "13 ## 7. N processors (parallel) (all)\n", "6 ## 8. Mindepth: min coverage for a cluster (s4,s5)\n", "4 ## 9. NQual: max # sites with qual < 20 (or see line 20)(s2)\n", ".85 ## 10. Wclust: clustering threshold as a decimal (s3,s6)\n", "rad ## 11. Datatype: rad,gbs,pairgbs,pairddrad,(others:see docs)(all)\n", "2 ## 12. MinCov: min samples in a final locus (s7)\n", "99 ## 13. MaxSH: max inds with shared hetero site (s7)\n", "c85d6m2p3H3N3 ## 14. Prefix name for final output (no spaces) (s7)\n", "==== optional params below this line =================================== affected step ==\n", " ## 15.opt.: select subset (prefix* only selector) (s2-s7)\n", " ## 16.opt.: add-on (outgroup) taxa (list or prefix*) (s6,s7)\n", " ## 17.opt.: exclude taxa (list or prefix*) (s7)\n", " ## 18.opt.: loc. of de-multiplexed data (s2)\n", "0 ## 19.opt.: maxM: N mismatches in barcodes (def= 1) (s1)\n", " ## 20.opt.: phred Qscore offset (def= 33) (s2)\n", " ## 21.opt.: filter: def=0=NQual 1=NQual+adapters. 2=strict (s2)\n", " ## 22.opt.: a priori E,H (def= 0.001,0.01, if not estimated) (s5)\n", "99 ## 23.opt.: maxN: max Ns in a cons seq (def=5) (s5)\n", "99 ## 24.opt.: maxH: max heterozyg. sites in cons seq (def=5) (s5)\n", " ## 25.opt.: ploidy: max alleles in cons seq (def=2;see docs) (s4,s5)\n", " ## 26.opt.: maxSNPs: (def=100). Paired (def=100,100) (s7)\n", "99 ## 27.opt.: maxIndels: within-clust,across-clust (def. 3,99) (s3,s7)\n", " ## 28.opt.: random number seed (def. 112233) (s3,s6,s7)\n", " ## 29.opt.: trim overhang left,right on final loci, def(0,0) (s7)\n", " ## 30.opt.: output formats: p,n,a,s,v,u,t,m,k,g,* (see docs) (s7)\n", " ## 31.opt.: maj. base call at depth>x0&0Y8-PM.[CU=?KJ]^7,&!UZM7$(?PC])/\\nM0LZ!\"8T 2J\\\\._7S]&M*R# 8[6$D(7U0>@IJ!5/*$\\\\6I16E&7?(E5S43)#0R%\\nMNV>A%))#?!$@EHGW0*4;J$>J<\\\\5X [!G_799#NK9<%Q0[\\'_:LQ!R5554,A@Z\\nMCJDM K *4C@P[]^]\\nM+HR3^PLRSH@9!X.S/N1+K/C[@I8(2Y?P\"HRPJ*K?PA;P\"[^&=Y!$S0$8SKU(\\nMDB@8.B;\"B_%AV#A\"&S153;OBG\"-LH;DI5,F &BR;\\\\5Q4N,FZ8*\\\\+35D/,AKL\\nMFEIJ5S6? OZ4S!\\\\-J:G0W3MC[MNAL@5Z?>KJ9BHW@:/ZAGS[*A\\nM5>U,AI<4[2*QU%+E\"_.\"9A1):>\"1E^HWGI(P_JP/9>-V3.[F9 \\\\3XQ\"W#*>HIA+7@%D\\\\&HY%ULTC\\nMC$#)4_HOSZ#D3ES9/=B3%ILR=H*^\\'B(#.!/6G(X>[N*\\'T>$\"+MH\"\\nMRC;$&3_9;@8,\\';%_<>/5GB>[;(%1F5^GF%Z5,!6U>>%EW_8*3#+&9QG$P3;P\\nM3I_9!DZB%K@NG,:_FUQICI7/F@-OT-)T;RP>J3B)6V#L7^B;*3BL*&M2#&+_\\nM_H8R6N--$T*2^:C_\\'\\\\!))R \\\\4*&T@(_DIBTU\"J,H)O@W)M!V3&PA&%ZH^P\"!\\nMQ[W Z8;C6Y\\\\!MVVHY$H:%/POO\\\\WX)1_\\'@,\\\\VP&VX^\"SYLYJ\\';90WS6)GDV/ \\nMG4_K4@FV\\\\M!-(]N\\'F+Q=A^RZZ?1@3S9%W]]^,U,/$D=1@%F&^8UJZ;XA[MLM\\nM-GI-FIQO@#]CLRP1VL6LD\"=-\"R(TU\\\\H8_Z\\'9(H247%X&Z[&A![OSG\\\\:!1%4@\\nM%]4C;YHB:Y\\'B.$EV!.T[VC\\'LSH*HTVHSCI1\\\\9HD6\\\\P(C1FZZCR!.\\\\,.(1($G\\nMY/DPOO=,N&V*MH_H06^>-&8&F#AM7,W1J.47<5I_)-,KJ_[@;67N\"U\\nM#86\\\\YIK7FS[!W,>VV[_+,%YS50MN_&[Q4>#.AC@]0R%PXL,16B#3VS-5PV\\\\6\\nM$1\"KXWAH%35/&ZPX>\\\\T0_C7@_U\"%8@2B3*HUL)_1>K [\\nM&W;COZ/;#7[N &L\\\\M9_JL=UH\\\\A8VDMM@_S6:8,MS7;;IY*=Y29%4:HR8RXI+\\n/NSNQ;&EX=U89_ >8=H2-\\n \\nend\\n'\n", "pyrad_params = pyrad_params.decode(\"uu\").decode(\"zlib\")\n", "print(pyrad_params)\n", "\n", "## If you want to make changes to the params file you can generate a new uuencoded string\n", "## by editing the pyrad/params-pyrad.txt file and running these lines. If you run this\n", "## at a terminal inside python shell then it'll print out nicely as one big fat line. \n", "#with open(os.path.join(PYRAD_DIR, \"params-pyrad.txt\")) as infile:\n", "# infile.read().encode(\"zlib\").encode(\"uu\")" ] }, { "cell_type": "code", "execution_count": 9, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time pyrad -p /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simno/paramsno-pyrad.txt -s 1234567\n", "\n", "\n", " ------------------------------------------------------------\n", " pyRAD : RADseq for phylogenetics & introgression analyses\n", " ------------------------------------------------------------\n", "\n", "\n", "\tstep 1: sorting reads by barcode\n", "\t .\tstep 2: editing raw reads \n", "\t............\n", "\tde-replicating files for clustering...\n", "\n", "\tstep 3: within-sample clustering of 12 samples at \n", "\t '.85' similarity. Running 12 parallel jobs\n", "\t \twith up to 6 threads per job. If needed, \n", "\t\tadjust to avoid CPU and MEM limits\n", "\n", "\tsample 1D_0 finished, 10000 loci\n", "\tsample 3I_0 finished, 10000 loci\n", "\tsample 3J_0 finished, 10000 loci\n", "\tsample 3K_0 finished, 10000 loci\n", "\tsample 1A_0 finished, 10000 loci\n", "\tsample 2F_0 finished, 10000 loci\n", "\tsample 1B_0 finished, 10000 loci\n", "\tsample 2E_0 finished, 10000 loci\n", "\tsample 2H_0 finished, 10000 loci\n", "\tsample 1C_0 finished, 10000 loci\n", "\tsample 3L_0 finished, 10000 loci\n", "\tsample 2G_0 finished, 10000 loci\n", "\n", "\tstep 4: estimating error rate and heterozygosity\n", "\t............\n", "\tstep 5: creating consensus seqs for 12 samples, using H=0.00198 E=0.00075\n", "\t............\n", "\tstep 6: clustering across 12 samples at '.85' similarity \n", "\n", "vsearch v2.0.3_linux_x86_64, 125.9GB RAM, 40 cores\n", "https://github.com/torognes/vsearch\n", "\n", "Reading file /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simno/clust.85/cat.haplos_ 100%\n", "10919996 nt in 120000 seqs, min 90, max 91, avg 91\n", "Counting unique k-mers 100%\n", "Clustering 100%\n", "Sorting clusters 100%\n", "Writing clusters 100%\n", "Clusters: 10000 Size min 12, max 12, avg 12.0\n", "Singletons: 0, 0.0% of seqs, 0.0% of clusters\n", "\n", "\tfinished clustering\n", "\tingroup 1A_0,1B_0,1C_0,1D_0,2E_0,2F_0,2G_0,2H_0,3I_0,3J_0,3K_0,3L_0\n", "\taddon \n", "\texclude \n", "\t.............\n", "\tfinal stats written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simno/stats/c85d6m2p3H3N3.stats\n", "\toutput files being written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simno/outfiles/ directory\n", "\n", "\n", "real\t11m6.442s\n", "user\t105m7.340s\n", "sys\t12m15.424s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time pyrad -p /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simlo/paramslo-pyrad.txt -s 1234567\n", "\n", "\n", " ------------------------------------------------------------\n", " pyRAD : RADseq for phylogenetics & introgression analyses\n", " ------------------------------------------------------------\n", "\n", "\n", "\tstep 1: sorting reads by barcode\n", "\t .\tstep 2: editing raw reads \n", "\t............\n", "\tde-replicating files for clustering...\n", "\n", "\tstep 3: within-sample clustering of 12 samples at \n", "\t '.85' similarity. Running 12 parallel jobs\n", "\t \twith up to 6 threads per job. If needed, \n", "\t\tadjust to avoid CPU and MEM limits\n", "\n", "\tsample 1D_0 finished, 10000 loci\n", "\tsample 2E_0 finished, 10000 loci\n", "\tsample 2H_0 finished, 10000 loci\n", "\tsample 1A_0 finished, 10000 loci\n", "\tsample 1C_0 finished, 10000 loci\n", "\tsample 1B_0 finished, 10000 loci\n", "\tsample 3L_0 finished, 10000 loci\n", "\tsample 2G_0 finished, 10000 loci\n", "\tsample 2F_0 finished, 10000 loci\n", "\tsample 3K_0 finished, 10000 loci\n", "\tsample 3J_0 finished, 10000 loci\n", "\tsample 3I_0 finished, 10000 loci\n", "\n", "\tstep 4: estimating error rate and heterozygosity\n", "\t............\n", "\tstep 5: creating consensus seqs for 12 samples, using H=0.00195 E=0.00075\n", "\t............\n", "\tstep 6: clustering across 12 samples at '.85' similarity \n", "\n", "vsearch v2.0.3_linux_x86_64, 125.9GB RAM, 40 cores\n", "https://github.com/torognes/vsearch\n", "\n", "Reading file /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simlo/clust.85/cat.haplos_ 100%\n", "10920527 nt in 120000 seqs, min 90, max 94, avg 91\n", "Counting unique k-mers 100%\n", "Clustering 100%\n", "Sorting clusters 100%\n", "Writing clusters 100%\n", "Clusters: 10000 Size min 12, max 12, avg 12.0\n", "Singletons: 0, 0.0% of seqs, 0.0% of clusters\n", "\n", "\tfinished clustering\n", "\tingroup 1A_0,1B_0,1C_0,1D_0,2E_0,2F_0,2G_0,2H_0,3I_0,3J_0,3K_0,3L_0\n", "\taddon \n", "\texclude \n", "\t.............\n", "\tfinal stats written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simlo/stats/c85d6m2p3H3N3.stats\n", "\toutput files being written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simlo/outfiles/ directory\n", "\n", "\n", "real\t11m2.628s\n", "user\t104m0.546s\n", "sys\t12m28.283s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time pyrad -p /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simhi/paramshi-pyrad.txt -s 1234567\n", "\n", "\n", " ------------------------------------------------------------\n", " pyRAD : RADseq for phylogenetics & introgression analyses\n", " ------------------------------------------------------------\n", "\n", "\n", "\tstep 1: sorting reads by barcode\n", "\t .\tstep 2: editing raw reads \n", "\t............\n", "\tde-replicating files for clustering...\n", "\n", "\tstep 3: within-sample clustering of 12 samples at \n", "\t '.85' similarity. Running 12 parallel jobs\n", "\t \twith up to 6 threads per job. If needed, \n", "\t\tadjust to avoid CPU and MEM limits\n", "\n", "\tsample 3I_0 finished, 10000 loci\n", "\tsample 1D_0 finished, 10000 loci\n", "\tsample 2H_0 finished, 10000 loci\n", "\tsample 2G_0 finished, 10000 loci\n", "\tsample 1A_0 finished, 10000 loci\n", "\tsample 2E_0 finished, 10000 loci\n", "\tsample 1B_0 finished, 10000 loci\n", "\tsample 3K_0 finished, 10000 loci\n", "\tsample 3J_0 finished, 10000 loci\n", "\tsample 2F_0 finished, 10001 loci\n", "\tsample 3L_0 finished, 10000 loci\n", "\tsample 1C_0 finished, 10000 loci\n", "\n", "\tstep 4: estimating error rate and heterozygosity\n", "\t............\n", "\tstep 5: creating consensus seqs for 12 samples, using H=0.00191 E=0.00075\n", "\t............\n", "\tstep 6: clustering across 12 samples at '.85' similarity \n", "\n", "vsearch v2.0.3_linux_x86_64, 125.9GB RAM, 40 cores\n", "https://github.com/torognes/vsearch\n", "\n", "Reading file /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simhi/clust.85/cat.haplos_ 100%\n", "10921211 nt in 120000 seqs, min 90, max 94, avg 91\n", "Counting unique k-mers 100%\n", "Clustering 100%\n", "Sorting clusters 100%\n", "Writing clusters 100%\n", "Clusters: 10002 Size min 1, max 12, avg 12.0\n", "Singletons: 1, 0.0% of seqs, 0.0% of clusters\n", "\n", "\tfinished clustering\n", "\tingroup 1A_0,1B_0,1C_0,1D_0,2E_0,2F_0,2G_0,2H_0,3I_0,3J_0,3K_0,3L_0\n", "\taddon \n", "\texclude \n", "\t.............\n", "\tfinal stats written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simhi/stats/c85d6m2p3H3N3.stats\n", "\toutput files being written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simhi/outfiles/ directory\n", "\n", "\n", "real\t11m5.615s\n", "user\t103m53.529s\n", "sys\t12m30.926s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time pyrad -p /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simlarge/paramslarge-pyrad.txt -s 1234567\n", "\n", "\n", " ------------------------------------------------------------\n", " pyRAD : RADseq for phylogenetics & introgression analyses\n", " ------------------------------------------------------------\n", "\n", "\n", "\tstep 1: sorting reads by barcode\n", "\t .\tstep 2: editing raw reads \n", "\t............\n", "\tde-replicating files for clustering...\n", "\n", "\tstep 3: within-sample clustering of 12 samples at \n", "\t '.85' similarity. Running 12 parallel jobs\n", "\t \twith up to 6 threads per job. If needed, \n", "\t\tadjust to avoid CPU and MEM limits\n", "\n", "\tsample 2G_0 finished, 96973 loci\n", "\tsample 3J_0 finished, 96941 loci\n", "\tsample 3I_0 finished, 96934 loci\n", "\tsample 3K_0 finished, 96942 loci\n", "\tsample 2H_0 finished, 96978 loci\n", "\tsample 1D_0 finished, 97002 loci\n", "\tsample 1C_0 finished, 96950 loci\n", "\tsample 1A_0 finished, 96943 loci\n", "\tsample 2E_0 finished, 96985 loci\n", "\tsample 2F_0 finished, 96943 loci\n", "\tsample 3L_0 finished, 96964 loci\n", "\tsample 1B_0 finished, 96980 loci\n", "\n", "\tstep 4: estimating error rate and heterozygosity\n", "\t............\n", "\tstep 5: creating consensus seqs for 12 samples, using H=0.00197 E=0.00075\n", "\t............\n", "\tstep 6: clustering across 12 samples at '.85' similarity \n", "\n", "vsearch v2.0.3_linux_x86_64, 125.9GB RAM, 40 cores\n", "https://github.com/torognes/vsearch\n", "\n", "Reading file /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simlarge/clust.85/cat.haplos_ 100%\n", "105866781 nt in 1163372 seqs, min 90, max 91, avg 91\n", "Counting unique k-mers 100%\n", "Clustering 100%\n", "Sorting clusters 100%\n", "Writing clusters 100%\n", "Clusters: 99999 Size min 1, max 12, avg 11.6\n", "Singletons: 48, 0.0% of seqs, 0.0% of clusters\n", "\n", "\tfinished clustering\n", "\tingroup 1A_0,1B_0,1C_0,1D_0,2E_0,2F_0,2G_0,2H_0,3I_0,3J_0,3K_0,3L_0\n", "\taddon \n", "\texclude \n", "\t.............\n", "\tfinal stats written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simlarge/stats/c85d6m2p3H3N3.stats\n", "\toutput files being written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/SIMDATA/simlarge/outfiles/ directory\n", "\n", "\n", "real\t92m41.293s\n", "user\t815m59.288s\n", "sys\t164m25.130s\n", "\n" ] } ], "source": [ "import subprocess\n", "\n", "## Set up directory structures. change the force flag if you want to\n", "## blow everything away and restart\n", "# force = True\n", "force = \"\"\n", "PYRAD_SIMOUT=os.path.join(PYRAD_DIR, \"SIMDATA/\")\n", "if force and os.path.exists(PYRAD_SIMOUT):\n", " shutil.rmtree(PYRAD_SIMOUT)\n", "if not os.path.exists(PYRAD_SIMOUT):\n", " os.makedirs(PYRAD_SIMOUT)\n", "for outdir in [\"no/\", \"lo/\", \"hi/\", \"large/\"]:\n", " tmpdir = os.path.join(PYRAD_SIMOUT, \"sim\"+outdir)\n", " if not os.path.exists(tmpdir):\n", " os.makedirs(tmpdir)\n", "\n", "## go to the pyrad simulated data output directory\n", "for dir in [\"no\", \"lo\", \"hi\", \"large\"]:\n", " ## Set barcode and raw data files\n", " simdata_dir = os.path.join(SIMULATION_DATA_DIR, \"sim\"+dir)\n", " bcodes = os.path.join(simdata_dir, \"sim{}_barcodes.txt\".format(dir))\n", " fastqs = os.path.join(simdata_dir, \"sim{}_R1_.fastq.gz\".format(dir))\n", " \n", " ## Set output directory and params file\n", " simout_dir = os.path.join(PYRAD_SIMOUT, \"sim\"+dir)\n", " my_params_file = os.path.join(simout_dir, \"params{}-pyrad.txt\".format(dir))\n", " \n", " ## Grab the generic pyrad params file and modify it for this run\n", " my_params = pyrad_params.replace(\"\", simout_dir)\n", " my_params = my_params.replace(\"\", fastqs)\n", " my_params = my_params.replace(\"\", bcodes)\n", "\n", " ## Write the params out to a file\n", " with open(my_params_file, 'w') as outfile:\n", " outfile.write(my_params)\n", "\n", " cmd = \"export PATH={workdir}/miniconda/bin:$PATH; \".format(workdir=WORK_DIR)\n", " cmd += \"time pyrad -p {params_file} -s 1234567\".format(params_file=my_params_file)\n", "\n", " ## Good sub for testing\n", " #cmd += \"time pyrad --version\"\n", "\n", " print(cmd)\n", " output = subprocess.check_output(cmd, shell=True, stderr=subprocess.STDOUT)\n", " print(output)" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## stacks simulated data assembly\n", "Output for stacks will be a bit trickier bcz we'll run gapped, ungapped and default on all sim data\n", "```\n", "WORK_DIR\n", " |____stacks\n", " |___SIMOUT\n", " |___gapped\n", " |____simno\n", " |____simlo\n", " |____simhi\n", " |____ungapped\n", " |\n", " |____default\n", "```\n", "\n", "We'll make calls to stacks that will look kind of like this:\n", "```\n", "* process_radtags args\n", " * -f input file\n", " * -i gzfastq\n", " * -b barcodes file\n", " * -e pstI\n", " * -o output dir\n", "\n", "process_radtags -f simno_R1_.fastq.gz -i gzfastq -b simno_barcodes.txt -e pstI -o simout\n", "\n", "* denovo_map.pl args\n", " * -m 2: Minimum depth to make a stack\n", " * -M 10: Number of mismatches per locus within individuals\n", " * -N 10: Number of allowed mismatches in secondary reads\n", " * -n 10: Number of mismatches when clustering across individuals \n", " \n", " Housekeeping flags:\n", " * -T 40: Number of threads\n", " * -b 1: Batch id (Can be anything)\n", " * -S: Disable sql\n", "\n", "denovo_map.pl -m 2 -M 10 -N 10 -n 10 -T 40 -b 1 -S --gapped -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -X 'populations:-m 6' -X 'ustacks:--max_locus_stacks 2' -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/ -s \n", "\n", "Times:\n", "\n", " * Demux small - 15 seconds\n", " * Demux large - 2m30s\n", " * denovo_map small - \n", " * denovo_map large - 23m" ] }, { "cell_type": "code", "execution_count": 335, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "##################################\n", "Doing - /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/ - simno\n", "##################################\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time process_radtags -f /home/iovercast/manuscript-analysis/simulated_data/simno/simno_R1_.fastq.gz -i gzfastq -b /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/stacks_barcodes.txt -e pstI -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno; time denovo_map.pl -T 40 -b 1 -S -m 2 -M 10 -N 10 -n 10 --gapped -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno --samples /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -O /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/stacks_popmap.txt\n", "Processing single-end data.\n", "Using Phred+33 encoding for quality scores.\n", "Found 1 input file(s).\n", "Searching for single-end, inlined barcodes.\n", "Loaded 12 barcodes (9bp).\n", "Processing file 1 of 1 [simno_R1_.fastq.gz]\n", " 2398782 total reads; -0 ambiguous barcodes; -0 ambiguous RAD-Tags; +0 recovered; -0 low quality reads; 2398782 retained reads.\n", "Closing files, flushing buffers...\n", "Outputing details to log: '/home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/process_radtags.log'\n", "\n", "2398782 total sequences;\n", " 0 ambiguous barcode drops;\n", " 0 low quality read drops;\n", " 0 ambiguous RAD-Tag drops;\n", "2398782 retained reads.\n", "\n", "real\t0m14.880s\n", "user\t0m14.727s\n", "sys\t0m0.024s\n", "Parsed population map: 12 files in 1 population and 1 group.\n", "Found 12 sample file(s).\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1A_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 1 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 1 of 12 [1A_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1B_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 2 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 2 of 12 [1B_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1C_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 3 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 3 of 12 [1C_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1D_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 4 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 4 of 12 [1D_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2E_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 5 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 5 of 12 [2E_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2F_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 6 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 6 of 12 [2F_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2G_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 7 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 7 of 12 [2G_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2H_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 8 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 8 of 12 [2H_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3I_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 9 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 9 of 12 [3I_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3J_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 10 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 10 of 12 [3J_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3K_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 11 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 11 of 12 [3K_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3L_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -i 12 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 12 of 12 [3L_0]\n", "\n", "Depths of Coverage for Processed Samples:\n", "1A_0: 19.953x\n", "1B_0: 19.9645x\n", "1C_0: 19.9678x\n", "1D_0: 20.018x\n", "2E_0: 19.9596x\n", "2F_0: 19.9424x\n", "2G_0: 20.0055x\n", "2H_0: 19.989x\n", "3I_0: 19.9392x\n", "3J_0: 19.9353x\n", "3K_0: 19.9284x\n", "3L_0: 19.9928x\n", "\n", "Generating catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3L_0 -p 40 -n 10 --gapped 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/batch_1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/3L_0 -p 40 --gapped 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/manuscript-analysis/miniconda/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -M /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simno/stacks_popmap.txt 2>&1\n", "\n", "real\t2m13.996s\n", "user\t12m57.180s\n", "sys\t0m5.520s\n", "\n", "##################################\n", "Doing - /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/ - simlo\n", "##################################\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time process_radtags -f /home/iovercast/manuscript-analysis/simulated_data/simlo/simlo_R1_.fastq.gz -i gzfastq -b /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/stacks_barcodes.txt -e pstI -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo; time denovo_map.pl -T 40 -b 1 -S -m 2 -M 10 -N 10 -n 10 --gapped -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo --samples /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -O /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/stacks_popmap.txt\n", "Processing single-end data.\n", "Using Phred+33 encoding for quality scores.\n", "Found 1 input file(s).\n", "Searching for single-end, inlined barcodes.\n", "Loaded 12 barcodes (9bp).\n", "Processing file 1 of 1 [simlo_R1_.fastq.gz]\n", " 2397592 total reads; -0 ambiguous barcodes; -0 ambiguous RAD-Tags; +0 recovered; -0 low quality reads; 2397592 retained reads.\n", "Closing files, flushing buffers...\n", "Outputing details to log: '/home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/process_radtags.log'\n", "\n", "2397592 total sequences;\n", " 0 ambiguous barcode drops;\n", " 0 low quality read drops;\n", " 0 ambiguous RAD-Tag drops;\n", "2397592 retained reads.\n", "\n", "real\t0m14.630s\n", "user\t0m14.476s\n", "sys\t0m0.023s\n", "Parsed population map: 12 files in 1 population and 1 group.\n", "Found 12 sample file(s).\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1A_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 1 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 1 of 12 [1A_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1B_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 2 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 2 of 12 [1B_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1C_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 3 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 3 of 12 [1C_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1D_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 4 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 4 of 12 [1D_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2E_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 5 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 5 of 12 [2E_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2F_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 6 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 6 of 12 [2F_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2G_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 7 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 7 of 12 [2G_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2H_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 8 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 8 of 12 [2H_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3I_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 9 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 9 of 12 [3I_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3J_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 10 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 10 of 12 [3J_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3K_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 11 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 11 of 12 [3K_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3L_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -i 12 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 12 of 12 [3L_0]\n", "\n", "Depths of Coverage for Processed Samples:\n", "1A_0: 19.9639x\n", "1B_0: 19.9531x\n", "1C_0: 19.9836x\n", "1D_0: 19.9599x\n", "2E_0: 19.9959x\n", "2F_0: 19.9282x\n", "2G_0: 19.9601x\n", "2H_0: 19.9246x\n", "3I_0: 19.9341x\n", "3J_0: 19.9345x\n", "3K_0: 19.9455x\n", "3L_0: 19.9836x\n", "\n", "Generating catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3L_0 -p 40 -n 10 --gapped 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/batch_1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/3L_0 -p 40 --gapped 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/manuscript-analysis/miniconda/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -M /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simlo/stacks_popmap.txt 2>&1\n", "\n", "real\t1m23.827s\n", "user\t10m11.612s\n", "sys\t0m4.204s\n", "\n", "##################################\n", "Doing - /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/ - simhi\n", "##################################\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time process_radtags -f /home/iovercast/manuscript-analysis/simulated_data/simhi/simhi_R1_.fastq.gz -i gzfastq -b /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/stacks_barcodes.txt -e pstI -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi; time denovo_map.pl -T 40 -b 1 -S -m 2 -M 10 -N 10 -n 10 --gapped -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi --samples /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -O /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/stacks_popmap.txt\n", "Processing single-end data.\n", "Using Phred+33 encoding for quality scores.\n", "Found 1 input file(s).\n", "Searching for single-end, inlined barcodes.\n", "Loaded 12 barcodes (9bp).\n", "Processing file 1 of 1 [simhi_R1_.fastq.gz]\n", " 2400054 total reads; -0 ambiguous barcodes; -0 ambiguous RAD-Tags; +0 recovered; -0 low quality reads; 2400054 retained reads.\n", "Closing files, flushing buffers...\n", "Outputing details to log: '/home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/process_radtags.log'\n", "\n", "2400054 total sequences;\n", " 0 ambiguous barcode drops;\n", " 0 low quality read drops;\n", " 0 ambiguous RAD-Tag drops;\n", "2400054 retained reads.\n", "\n", "real\t0m14.621s\n", "user\t0m14.574s\n", "sys\t0m0.025s\n", "Parsed population map: 12 files in 1 population and 1 group.\n", "Found 12 sample file(s).\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1A_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 1 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 1 of 12 [1A_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1B_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 2 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 2 of 12 [1B_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1C_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 3 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 3 of 12 [1C_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1D_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 4 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 4 of 12 [1D_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2E_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 5 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 5 of 12 [2E_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2F_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 6 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 6 of 12 [2F_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2G_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 7 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 7 of 12 [2G_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2H_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 8 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 8 of 12 [2H_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3I_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 9 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 9 of 12 [3I_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3J_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 10 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 10 of 12 [3J_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3K_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 11 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 11 of 12 [3K_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3L_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -i 12 -r -m 2 -p 40 -M 10 -N 10 --gapped 2>&1\n", "Identifying unique stacks; file 12 of 12 [3L_0]\n", "\n", "Depths of Coverage for Processed Samples:\n", "1A_0: 19.9321x\n", "1B_0: 19.9699x\n", "1C_0: 19.9865x\n", "1D_0: 20.0289x\n", "2E_0: 19.9526x\n", "2F_0: 19.9634x\n", "2G_0: 19.9455x\n", "2H_0: 19.9461x\n", "3I_0: 19.9013x\n", "3J_0: 19.9012x\n", "3K_0: 19.938x\n", "3L_0: 19.9768x\n", "\n", "Generating catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3L_0 -p 40 -n 10 --gapped 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/batch_1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/3L_0 -p 40 --gapped 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/manuscript-analysis/miniconda/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -M /home/iovercast/manuscript-analysis/stacks/SIMDATA/gapped/simhi/stacks_popmap.txt 2>&1\n", "\n", "real\t1m53.167s\n", "user\t12m41.126s\n", "sys\t0m4.751s\n", "\n", "##################################\n", "Doing - /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/ - simno\n", "##################################\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time process_radtags -f /home/iovercast/manuscript-analysis/simulated_data/simno/simno_R1_.fastq.gz -i gzfastq -b /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/stacks_barcodes.txt -e pstI -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno; time denovo_map.pl -T 40 -b 1 -S -m 2 -M 10 -N 10 -n 10 -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno --samples /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -O /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/stacks_popmap.txt\n", "Processing single-end data.\n", "Using Phred+33 encoding for quality scores.\n", "Found 1 input file(s).\n", "Searching for single-end, inlined barcodes.\n", "Loaded 12 barcodes (9bp).\n", "Processing file 1 of 1 [simno_R1_.fastq.gz]\n", " 2398782 total reads; -0 ambiguous barcodes; -0 ambiguous RAD-Tags; +0 recovered; -0 low quality reads; 2398782 retained reads.\n", "Closing files, flushing buffers...\n", "Outputing details to log: '/home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/process_radtags.log'\n", "\n", "2398782 total sequences;\n", " 0 ambiguous barcode drops;\n", " 0 low quality read drops;\n", " 0 ambiguous RAD-Tag drops;\n", "2398782 retained reads.\n", "\n", "real\t0m14.541s\n", "user\t0m14.493s\n", "sys\t0m0.014s\n", "Parsed population map: 12 files in 1 population and 1 group.\n", "Found 12 sample file(s).\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1A_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 1 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 1 of 12 [1A_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1B_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 2 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 2 of 12 [1B_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1C_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 3 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 3 of 12 [1C_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1D_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 4 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 4 of 12 [1D_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2E_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 5 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 5 of 12 [2E_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2F_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 6 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 6 of 12 [2F_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2G_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 7 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 7 of 12 [2G_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2H_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 8 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 8 of 12 [2H_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3I_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 9 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 9 of 12 [3I_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3J_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 10 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 10 of 12 [3J_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3K_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 11 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 11 of 12 [3K_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3L_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -i 12 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 12 of 12 [3L_0]\n", "\n", "Depths of Coverage for Processed Samples:\n", "1A_0: 19.953x\n", "1B_0: 19.9645x\n", "1C_0: 19.9678x\n", "1D_0: 20.018x\n", "2E_0: 19.9596x\n", "2F_0: 19.9424x\n", "2G_0: 20.0055x\n", "2H_0: 19.989x\n", "3I_0: 19.9392x\n", "3J_0: 19.9353x\n", "3K_0: 19.9284x\n", "3L_0: 19.9928x\n", "\n", "Generating catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3L_0 -p 40 -n 10 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/batch_1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/3L_0 -p 40 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/manuscript-analysis/miniconda/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -M /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simno/stacks_popmap.txt 2>&1\n", "\n", "real\t1m47.509s\n", "user\t12m10.832s\n", "sys\t0m4.458s\n", "\n", "##################################\n", "Doing - /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/ - simlo\n", "##################################\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time process_radtags -f /home/iovercast/manuscript-analysis/simulated_data/simlo/simlo_R1_.fastq.gz -i gzfastq -b /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/stacks_barcodes.txt -e pstI -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo; time denovo_map.pl -T 40 -b 1 -S -m 2 -M 10 -N 10 -n 10 -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo --samples /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -O /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/stacks_popmap.txt\n", "Processing single-end data.\n", "Using Phred+33 encoding for quality scores.\n", "Found 1 input file(s).\n", "Searching for single-end, inlined barcodes.\n", "Loaded 12 barcodes (9bp).\n", "Processing file 1 of 1 [simlo_R1_.fastq.gz]\n", " 2397592 total reads; -0 ambiguous barcodes; -0 ambiguous RAD-Tags; +0 recovered; -0 low quality reads; 2397592 retained reads.\n", "Closing files, flushing buffers...\n", "Outputing details to log: '/home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/process_radtags.log'\n", "\n", "2397592 total sequences;\n", " 0 ambiguous barcode drops;\n", " 0 low quality read drops;\n", " 0 ambiguous RAD-Tag drops;\n", "2397592 retained reads.\n", "\n", "real\t0m14.472s\n", "user\t0m14.309s\n", "sys\t0m0.020s\n", "Parsed population map: 12 files in 1 population and 1 group.\n", "Found 12 sample file(s).\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1A_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 1 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 1 of 12 [1A_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1B_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 2 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 2 of 12 [1B_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1C_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 3 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 3 of 12 [1C_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1D_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 4 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 4 of 12 [1D_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2E_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 5 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 5 of 12 [2E_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2F_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 6 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 6 of 12 [2F_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2G_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 7 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 7 of 12 [2G_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2H_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 8 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 8 of 12 [2H_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3I_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 9 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 9 of 12 [3I_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3J_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 10 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 10 of 12 [3J_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3K_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 11 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 11 of 12 [3K_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3L_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -i 12 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 12 of 12 [3L_0]\n", "\n", "Depths of Coverage for Processed Samples:\n", "1A_0: 19.924x\n", "1B_0: 19.9052x\n", "1C_0: 19.9236x\n", "1D_0: 19.906x\n", "2E_0: 19.9419x\n", "2F_0: 19.8546x\n", "2G_0: 19.8844x\n", "2H_0: 19.8748x\n", "3I_0: 19.8683x\n", "3J_0: 19.8589x\n", "3K_0: 19.8598x\n", "3L_0: 19.9236x\n", "\n", "Generating catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3L_0 -p 40 -n 10 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/batch_1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/3L_0 -p 40 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/manuscript-analysis/miniconda/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -M /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simlo/stacks_popmap.txt 2>&1\n", "\n", "real\t1m49.086s\n", "user\t12m2.639s\n", "sys\t0m4.565s\n", "\n", "##################################\n", "Doing - /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/ - simhi\n", "##################################\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time process_radtags -f /home/iovercast/manuscript-analysis/simulated_data/simhi/simhi_R1_.fastq.gz -i gzfastq -b /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/stacks_barcodes.txt -e pstI -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi; time denovo_map.pl -T 40 -b 1 -S -m 2 -M 10 -N 10 -n 10 -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi --samples /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -O /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/stacks_popmap.txt\n", "Processing single-end data.\n", "Using Phred+33 encoding for quality scores.\n", "Found 1 input file(s).\n", "Searching for single-end, inlined barcodes.\n", "Loaded 12 barcodes (9bp).\n", "Processing file 1 of 1 [simhi_R1_.fastq.gz]\n", " 2400054 total reads; -0 ambiguous barcodes; -0 ambiguous RAD-Tags; +0 recovered; -0 low quality reads; 2400054 retained reads.\n", "Closing files, flushing buffers...\n", "Outputing details to log: '/home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/process_radtags.log'\n", "\n", "2400054 total sequences;\n", " 0 ambiguous barcode drops;\n", " 0 low quality read drops;\n", " 0 ambiguous RAD-Tag drops;\n", "2400054 retained reads.\n", "\n", "real\t0m16.147s\n", "user\t0m15.936s\n", "sys\t0m0.031s\n", "Parsed population map: 12 files in 1 population and 1 group.\n", "Found 12 sample file(s).\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1A_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 1 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 1 of 12 [1A_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1B_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 2 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 2 of 12 [1B_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1C_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 3 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 3 of 12 [1C_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1D_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 4 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 4 of 12 [1D_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2E_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 5 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 5 of 12 [2E_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2F_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 6 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 6 of 12 [2F_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2G_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 7 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 7 of 12 [2G_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2H_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 8 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 8 of 12 [2H_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3I_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 9 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 9 of 12 [3I_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3J_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 10 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 10 of 12 [3J_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3K_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 11 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 11 of 12 [3K_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3L_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -i 12 -r -m 2 -p 40 -M 10 -N 10 2>&1\n", "Identifying unique stacks; file 12 of 12 [3L_0]\n", "\n", "Depths of Coverage for Processed Samples:\n", "1A_0: 19.8407x\n", "1B_0: 19.8289x\n", "1C_0: 19.9126x\n", "1D_0: 19.9053x\n", "2E_0: 19.8216x\n", "2F_0: 19.8679x\n", "2G_0: 19.8422x\n", "2H_0: 19.8448x\n", "3I_0: 19.7883x\n", "3J_0: 19.7765x\n", "3K_0: 19.8268x\n", "3L_0: 19.8712x\n", "\n", "Generating catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3L_0 -p 40 -n 10 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/batch_1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/3L_0 -p 40 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/manuscript-analysis/miniconda/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -M /home/iovercast/manuscript-analysis/stacks/SIMDATA/ungapped/simhi/stacks_popmap.txt 2>&1\n", "\n", "real\t1m53.780s\n", "user\t12m1.654s\n", "sys\t0m5.007s\n", "\n", "##################################\n", "Doing - /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/ - simno\n", "##################################\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time process_radtags -f /home/iovercast/manuscript-analysis/simulated_data/simno/simno_R1_.fastq.gz -i gzfastq -b /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/stacks_barcodes.txt -e pstI -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno; time denovo_map.pl -T 40 -b 1 -S -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno --samples /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -O /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/stacks_popmap.txt\n", "Processing single-end data.\n", "Using Phred+33 encoding for quality scores.\n", "Found 1 input file(s).\n", "Searching for single-end, inlined barcodes.\n", "Loaded 12 barcodes (9bp).\n", "Processing file 1 of 1 [simno_R1_.fastq.gz]\n", " 2398782 total reads; -0 ambiguous barcodes; -0 ambiguous RAD-Tags; +0 recovered; -0 low quality reads; 2398782 retained reads.\n", "Closing files, flushing buffers...\n", "Outputing details to log: '/home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/process_radtags.log'\n", "\n", "2398782 total sequences;\n", " 0 ambiguous barcode drops;\n", " 0 low quality read drops;\n", " 0 ambiguous RAD-Tag drops;\n", "2398782 retained reads.\n", "\n", "real\t0m14.677s\n", "user\t0m14.512s\n", "sys\t0m0.025s\n", "Parsed population map: 12 files in 1 population and 1 group.\n", "Found 12 sample file(s).\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1A_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 1 -r -p 40 2>&1\n", "Identifying unique stacks; file 1 of 12 [1A_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1B_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 2 -r -p 40 2>&1\n", "Identifying unique stacks; file 2 of 12 [1B_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1C_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 3 -r -p 40 2>&1\n", "Identifying unique stacks; file 3 of 12 [1C_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1D_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 4 -r -p 40 2>&1\n", "Identifying unique stacks; file 4 of 12 [1D_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2E_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 5 -r -p 40 2>&1\n", "Identifying unique stacks; file 5 of 12 [2E_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2F_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 6 -r -p 40 2>&1\n", "Identifying unique stacks; file 6 of 12 [2F_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2G_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 7 -r -p 40 2>&1\n", "Identifying unique stacks; file 7 of 12 [2G_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2H_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 8 -r -p 40 2>&1\n", "Identifying unique stacks; file 8 of 12 [2H_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3I_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 9 -r -p 40 2>&1\n", "Identifying unique stacks; file 9 of 12 [3I_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3J_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 10 -r -p 40 2>&1\n", "Identifying unique stacks; file 10 of 12 [3J_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3K_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 11 -r -p 40 2>&1\n", "Identifying unique stacks; file 11 of 12 [3K_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3L_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -i 12 -r -p 40 2>&1\n", "Identifying unique stacks; file 12 of 12 [3L_0]\n", "\n", "Depths of Coverage for Processed Samples:\n", "1A_0: 19.9051x\n", "1B_0: 19.9106x\n", "1C_0: 19.9199x\n", "1D_0: 19.9458x\n", "2E_0: 19.8817x\n", "2F_0: 19.8906x\n", "2G_0: 19.9295x\n", "2H_0: 19.9191x\n", "3I_0: 19.8735x\n", "3J_0: 19.8835x\n", "3K_0: 19.8506x\n", "3L_0: 19.9288x\n", "\n", "Generating catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3L_0 -p 40 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/batch_1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/3L_0 -p 40 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/manuscript-analysis/miniconda/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -M /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simno/stacks_popmap.txt 2>&1\n", "\n", "real\t1m52.769s\n", "user\t3m2.639s\n", "sys\t0m4.437s\n", "\n", "##################################\n", "Doing - /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/ - simlo\n", "##################################\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time process_radtags -f /home/iovercast/manuscript-analysis/simulated_data/simlo/simlo_R1_.fastq.gz -i gzfastq -b /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/stacks_barcodes.txt -e pstI -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo; time denovo_map.pl -T 40 -b 1 -S -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo --samples /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -O /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/stacks_popmap.txt\n", "Processing single-end data.\n", "Using Phred+33 encoding for quality scores.\n", "Found 1 input file(s).\n", "Searching for single-end, inlined barcodes.\n", "Loaded 12 barcodes (9bp).\n", "Processing file 1 of 1 [simlo_R1_.fastq.gz]\n", " 2397592 total reads; -0 ambiguous barcodes; -0 ambiguous RAD-Tags; +0 recovered; -0 low quality reads; 2397592 retained reads.\n", "Closing files, flushing buffers...\n", "Outputing details to log: '/home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/process_radtags.log'\n", "\n", "2397592 total sequences;\n", " 0 ambiguous barcode drops;\n", " 0 low quality read drops;\n", " 0 ambiguous RAD-Tag drops;\n", "2397592 retained reads.\n", "\n", "real\t0m15.063s\n", "user\t0m14.907s\n", "sys\t0m0.027s\n", "Parsed population map: 12 files in 1 population and 1 group.\n", "Found 12 sample file(s).\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1A_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 1 -r -p 40 2>&1\n", "Identifying unique stacks; file 1 of 12 [1A_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1B_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 2 -r -p 40 2>&1\n", "Identifying unique stacks; file 2 of 12 [1B_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1C_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 3 -r -p 40 2>&1\n", "Identifying unique stacks; file 3 of 12 [1C_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1D_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 4 -r -p 40 2>&1\n", "Identifying unique stacks; file 4 of 12 [1D_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2E_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 5 -r -p 40 2>&1\n", "Identifying unique stacks; file 5 of 12 [2E_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2F_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 6 -r -p 40 2>&1\n", "Identifying unique stacks; file 6 of 12 [2F_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2G_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 7 -r -p 40 2>&1\n", "Identifying unique stacks; file 7 of 12 [2G_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2H_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 8 -r -p 40 2>&1\n", "Identifying unique stacks; file 8 of 12 [2H_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3I_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 9 -r -p 40 2>&1\n", "Identifying unique stacks; file 9 of 12 [3I_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3J_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 10 -r -p 40 2>&1\n", "Identifying unique stacks; file 10 of 12 [3J_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3K_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 11 -r -p 40 2>&1\n", "Identifying unique stacks; file 11 of 12 [3K_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3L_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -i 12 -r -p 40 2>&1\n", "Identifying unique stacks; file 12 of 12 [3L_0]\n", "\n", "Depths of Coverage for Processed Samples:\n", "1A_0: 19.8703x\n", "1B_0: 19.8456x\n", "1C_0: 19.8661x\n", "1D_0: 19.8248x\n", "2E_0: 19.8685x\n", "2F_0: 19.7992x\n", "2G_0: 19.8052x\n", "2H_0: 19.8035x\n", "3I_0: 19.8049x\n", "3J_0: 19.8094x\n", "3K_0: 19.7808x\n", "3L_0: 19.8502x\n", "\n", "Generating catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3L_0 -p 40 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/batch_1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/3L_0 -p 40 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/manuscript-analysis/miniconda/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -M /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simlo/stacks_popmap.txt 2>&1\n", "\n", "real\t1m53.102s\n", "user\t3m3.499s\n", "sys\t0m5.124s\n", "\n", "##################################\n", "Doing - /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/ - simhi\n", "##################################\n", "export PATH=/home/iovercast/manuscript-analysis//miniconda/bin:$PATH; time process_radtags -f /home/iovercast/manuscript-analysis/simulated_data/simhi/simhi_R1_.fastq.gz -i gzfastq -b /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/stacks_barcodes.txt -e pstI -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi; time denovo_map.pl -T 40 -b 1 -S -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi --samples /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -O /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/stacks_popmap.txt\n", "Processing single-end data.\n", "Using Phred+33 encoding for quality scores.\n", "Found 1 input file(s).\n", "Searching for single-end, inlined barcodes.\n", "Loaded 12 barcodes (9bp).\n", "Processing file 1 of 1 [simhi_R1_.fastq.gz]\n", " 2400054 total reads; -0 ambiguous barcodes; -0 ambiguous RAD-Tags; +0 recovered; -0 low quality reads; 2400054 retained reads.\n", "Closing files, flushing buffers...\n", "Outputing details to log: '/home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/process_radtags.log'\n", "\n", "2400054 total sequences;\n", " 0 ambiguous barcode drops;\n", " 0 low quality read drops;\n", " 0 ambiguous RAD-Tag drops;\n", "2400054 retained reads.\n", "\n", "real\t0m14.688s\n", "user\t0m14.534s\n", "sys\t0m0.024s\n", "Parsed population map: 12 files in 1 population and 1 group.\n", "Found 12 sample file(s).\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1A_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 1 -r -p 40 2>&1\n", "Identifying unique stacks; file 1 of 12 [1A_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1B_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 2 -r -p 40 2>&1\n", "Identifying unique stacks; file 2 of 12 [1B_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1C_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 3 -r -p 40 2>&1\n", "Identifying unique stacks; file 3 of 12 [1C_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1D_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 4 -r -p 40 2>&1\n", "Identifying unique stacks; file 4 of 12 [1D_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2E_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 5 -r -p 40 2>&1\n", "Identifying unique stacks; file 5 of 12 [2E_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2F_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 6 -r -p 40 2>&1\n", "Identifying unique stacks; file 6 of 12 [2F_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2G_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 7 -r -p 40 2>&1\n", "Identifying unique stacks; file 7 of 12 [2G_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2H_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 8 -r -p 40 2>&1\n", "Identifying unique stacks; file 8 of 12 [2H_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3I_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 9 -r -p 40 2>&1\n", "Identifying unique stacks; file 9 of 12 [3I_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3J_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 10 -r -p 40 2>&1\n", "Identifying unique stacks; file 10 of 12 [3J_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3K_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 11 -r -p 40 2>&1\n", "Identifying unique stacks; file 11 of 12 [3K_0]\n", " /home/iovercast/manuscript-analysis/miniconda/bin/ustacks -t gzfastq -f /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3L_0.fq.gz -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -i 12 -r -p 40 2>&1\n", "Identifying unique stacks; file 12 of 12 [3L_0]\n", "\n", "Depths of Coverage for Processed Samples:\n", "1A_0: 19.7736x\n", "1B_0: 19.7582x\n", "1C_0: 19.8454x\n", "1D_0: 19.8244x\n", "2E_0: 19.7413x\n", "2F_0: 19.801x\n", "2G_0: 19.7575x\n", "2H_0: 19.7719x\n", "3I_0: 19.7156x\n", "3J_0: 19.7059x\n", "3K_0: 19.752x\n", "3L_0: 19.8041x\n", "\n", "Generating catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3L_0 -p 40 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/manuscript-analysis/miniconda/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/batch_1 -o /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1A_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1B_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1C_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/1D_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2E_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2F_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2G_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/2H_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3I_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3J_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3K_0 -s /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/3L_0 -p 40 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/manuscript-analysis/miniconda/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -M /home/iovercast/manuscript-analysis/stacks/SIMDATA/default/simhi/stacks_popmap.txt 2>&1\n", "\n", "real\t1m55.680s\n", "user\t3m6.456s\n", "sys\t0m5.915s\n", "\n" ] } ], "source": [ "## These are the python variables for the main stacks simout paths: \n", "## STACKS_GAP_SIMOUT, STACKS_UNGAP_SIMOUT, STACKS_DEFAULT_SIMOUT\n", "\n", "STACKS_SIMOUT=os.path.join(STACKS_DIR, \"SIMDATA/\")\n", "STACKS_GAP_SIMOUT=os.path.join(STACKS_SIMOUT, \"gapped/\")\n", "STACKS_UNGAP_SIMOUT=os.path.join(STACKS_SIMOUT, \"ungapped/\")\n", "STACKS_DEFAULT_SIMOUT=os.path.join(STACKS_SIMOUT, \"default/\")\n", "\n", "force = True\n", "#force = \"\"\n", "if force and os.path.exists(STACKS_SIMOUT):\n", " shutil.rmtree(STACKS_SIMOUT)\n", "if not os.path.exists(STACKS_SIMOUT):\n", " os.makedirs(STACKS_SIMOUT)\n", "\n", "## Make the sumulation output directories if they don't already exist\n", "for dir in [STACKS_GAP_SIMOUT, STACKS_UNGAP_SIMOUT, STACKS_DEFAULT_SIMOUT]:\n", " if not os.path.exists(dir):\n", " os.makedirs(dir)\n", " for outdir in [\"no/\", \"lo/\", \"hi/\", \"large/\"]:\n", " tmpdir = os.path.join(dir, \"sim\"+outdir)\n", " if not os.path.exists(tmpdir):\n", " os.makedirs(tmpdir)\n", "\n", "## Do the stacks sim analysis\n", "for dir in [STACKS_GAP_SIMOUT, STACKS_UNGAP_SIMOUT, STACKS_DEFAULT_SIMOUT]:\n", "\n", " for sim in [\"no\", \"lo\", \"hi\", \"large\"]:\n", " print(\"##################################\")\n", " print(\"Doing - \" + dir + \" - sim\" + sim)\n", " print(\"##################################\")\n", " ## Where simulation results will be written\n", " simout_dir = os.path.join(dir, \"sim\"+sim)\n", " \n", " ## Where the rawdata and barcodes live\n", " simdata_dir = os.path.join(SIMULATION_DATA_DIR, \"sim\"+sim)\n", " ## We can just use the sim barcodes file as the stacks population map\n", " ## because all it really is doing is reading the first column, which\n", " ## is the names of samples to use in denovo_map.pl\n", " ipyrad_bcodes = os.path.join(simdata_dir, \"sim{}_barcodes.txt\".format(sim))\n", " fastqs = os.path.join(simdata_dir, \"sim{}_R1_.fastq.gz\".format(sim))\n", "\n", " ## Munge the ipyrad barcodes file into the format stacks wants for the population map\n", " ## file (ie sample name then population). Here we just assign all samples to be from \n", " ## population 1\n", " popmap = os.path.join(simout_dir, \"stacks_popmap.txt\")\n", " with open(ipyrad_bcodes) as infile:\n", " with open(popmap, 'w') as outfile:\n", " lines = infile.readlines()\n", " for line in lines:\n", " ## This is just setting all samples to population 1 \n", " outfile.write(line.split()[0]+\"\\t1\\n\")\n", " \n", " ## Munge the sim barcodes file into the format that stacks wants\n", " ## basically the inverse of what ipyrad uses barcode first then sample name\n", " ## There is probably a fancier/smarter way to do this, but this works.\n", " bcodes = os.path.join(simout_dir, \"stacks_barcodes.txt\")\n", " with open(ipyrad_bcodes, 'r') as infile:\n", " with open(bcodes, 'w') as outfile:\n", " lines = infile.readlines()\n", " for line in lines:\n", " if line:\n", " outfile.write(\"\\t\".join(line.split()[::-1])+\"\\n\")\n", "\n", " ## Export the path so we can actually find the binaries\n", " cmd = \"export PATH={workdir}/miniconda/bin:$PATH; \".format(workdir=WORK_DIR)\n", "\n", " ## Add the command to process_radtags.\n", " ## Yes we don't have to do this over and over, but it's fast, and also ipyrad/pyrad\n", " ## are paying the same demultiplex penalty for every run.\n", " cmd += \"time process_radtags -f {simdata} -i gzfastq -b {barcodes} -e pstI -o {simout}; \"\\\n", " .format(simdata=fastqs, barcodes=bcodes, simout=simout_dir)\n", " \n", " ## Toggle the dryrun flag for testing\n", " #DRYRUN=\"-d\"\n", " DRYRUN=\"\"\n", "\n", " ## Don't add all the fancy stuff if you just want to do the default assembly\n", " ADDITIONAL_PARAMS = \"\"\n", " GAPPED = \"\"\n", " if not dir == STACKS_DEFAULT_SIMOUT:\n", " ADDITIONAL_PARAMS = \" -m 2 -M 10 -N 10 -n 10 \"\n", " if dir == STACKS_GAP_SIMOUT:\n", " ADDITIONAL_PARAMS += \" --gapped \"\n", " \n", " OUTPUT_FORMATS = \"--vcf --genepop --structure --phase --fastphase --phylip \"\n", " cmd += \"time denovo_map.pl -T 40 -b 1 -S \" + DRYRUN + ADDITIONAL_PARAMS\\\n", " + \" -X \\'populations:\" + OUTPUT_FORMATS + \"\\'\"\\\n", " + \" -o \" + simout_dir + \" --samples \" + simout_dir + \" -O \" + popmap \n", "\n", " print(cmd)\n", " output = subprocess.check_output(cmd, shell=True, stderr=subprocess.STDOUT)\n", " print(output)\n" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## aftrRAD simulated data assembly\n", "aftrRAD throws out any read with one or more bases less than \n", "the phred qscore `minQual` setting. You can't really tune how many \n", "low quality bases to retain. For the simulated data we are simulating\n", "all qscores arbitrarily high, so this isn't a problem. For\n", "empirical we might have to think about lowering minQual...\n", "\n", "Actually, throwing out many reads seems to be the aftrRAD strategy. I think\n", "its trying to reduce the number of potential reads to match (reduce singletons\n", "from error) because it \n", "\n", "Performance on 10000 100bp simulated dataset:\n", "* ~200 minutes\n", "\n", "Performance on 100000 100bp simulated dataset:\n", "* AftrRad.pl - ~4500 minutes\n", "* Genotype.pl - 13 days (my notebook crashed, but it finished in about this long)" ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": false, "scrolled": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "Doing - no\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl AftrRAD.pl re-TGCAG minDepth-6 P2-noP2 minIden-85 stringLength-99DataPath-Data/ BarcodePath-Barcodes/ maxProcesses-40\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "minParalog\t5\n", "minDepth\t6\n", "MaxH\t90\n", "P2\tnoP2\n", "BarcodePath\tBarcodes/\n", "minQual\t20\n", "re\tTGCAG\n", "numIndels\t3\n", "Phred\t33\n", "minIden\t85\n", "maxProcesses\t40\n", "stringLength\t99DataPath\n", "dplexedData\t0\n", "Help\t0\n", "DataPath\tData/\n", "\n", "\n", "Running AftrRAD...\n", "\n", "Recognized 1 data files.\n", "\n", "File names are...\n", " sim-no.txt \n", "\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "The barcodes you entered for data file 0.0 (file sim-no.txt) are...\n", " CATCATCAT TCAACTCTC AATACTAGT GAGTGACAG ATGGACCTT CGAGTTTCC TGTTAATAA TCCGTAGCC GTTTCAATC AGAATCTGC AATCGCCGT AGTCCGCTA \n", "\n", "The names you entered for data file 0.0 (file sim-no.txt) are...\n", " 1A_0 1B_0 1C_0 1D_0 2E_0 2F_0 2G_0 2H_0 3I_0 3J_0 3K_0 3L_0 \n", "\n", "\n", "Demultiplexing samples for data file sim-no.txt.\n", "\n", "Identifying unique sequences for each individual.\n", "23741 TempFiles/SubTemp_0/UniqueWithCountsIndividual1A_0.txt\n", "23790 TempFiles/SubTemp_0/UniqueWithCountsIndividual1B_0.txt\n", "23981 TempFiles/SubTemp_0/UniqueWithCountsIndividual1C_0.txt\n", "23890 TempFiles/SubTemp_0/UniqueWithCountsIndividual1D_0.txt\n", "23919 TempFiles/SubTemp_0/UniqueWithCountsIndividual2E_0.txt\n", "23925 TempFiles/SubTemp_0/UniqueWithCountsIndividual2F_0.txt\n", "24022 TempFiles/SubTemp_0/UniqueWithCountsIndividual2G_0.txt\n", "23860 TempFiles/SubTemp_0/UniqueWithCountsIndividual2H_0.txt\n", "23843 TempFiles/SubTemp_0/UniqueWithCountsIndividual3I_0.txt\n", "23709 TempFiles/SubTemp_0/UniqueWithCountsIndividual3J_0.txt\n", "23703 TempFiles/SubTemp_0/UniqueWithCountsIndividual3K_0.txt\n", "23986 TempFiles/SubTemp_0/UniqueWithCountsIndividual3L_0.txt\n", "\n", "Identifying all unique sequences within the dataset.\n", "\n", "Creating file to test mean read counts.\n", "Testing nonzero means in read counts to identify error reads.\n", "\n", "Performing heuristic search of all pairwise comparisons to identify potentially allelic pairs.\n", "Rapidly searching unique sequence number 1000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 2000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 3000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 4000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 5000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 6000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 7000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 8000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 9000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 10000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 11000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 12000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 13000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 14000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 15000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 16000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 17000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 18000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 19000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 20000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 21000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 22000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 23000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 24000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 25000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 26000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 27000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 28000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 29000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 30000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 31000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 32000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 33000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 34000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 35000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 36000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 37000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 38000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 39000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 40000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 41000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 42000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 43000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 44000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 45000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 46000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 47000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 48000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 49000 of 50695 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 50000 of 50695 for potentially allelic read pairs.\n", "\n", "Number of pairwise comparisons to align and evaluate is 117495\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 40000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "Working on alignment 80000 of 117495.\n", "\n", "Scoring each pairwise alignment. Alignments exceeding 85 % similarity will be retained.\n", "Parsing aligned sequences into candidate loci.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 4000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Working on alignment 8000 of 9958.\n", "Finished writing the mafft output.\n", "Identifying and removing paralogous loci.\n", "\n", "\n", "Loci have been identified. Aligned polymorphic loci are stored in the Output directory. Read counts for the alleles at these loci can be found for each individual in the files stored in TempFiles/RawReadCountFiles.\n", "\n", "Demultiplexing and other general information can be found for each dataset analyzed in the Output/RunInfo directory.\n", "\n", "Next, run Genotype.pl, followed by FilterSNPs.pl to genotype all individuals and output SNPs for subsequent analyses.\n", "\n", "\n", "real\t53m20.317s\n", "user\t57m14.741s\n", "sys\t30m25.360s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl Genotype.pl MinReads-6 subset-0 maxProcesses-40\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "subsetfile\tSamplesForSubset.txt\n", "pvalLow\t0.0001\n", "pvalThresh\t100\n", "subset\t0\n", "maxProcesses\t40\n", "MinReads\t6\n", "pvalHigh\t1e-05\n", "Help\t0\n", "\n", "\n", "Running Genotype.pl...\n", "\n", "Scoring genotypes at all nonparalogous loci.\n", "\n", "Currently genotyping sample...1C_0\t1B_0\t2F_0\t3L_0\t3I_0\t2G_0\t1A_0\t2H_0\t3K_0\t1D_0\t2E_0\t3J_0\tRunning R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "cat: TempFiles/SubGenotypes_12/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_13/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_14/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_15/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_16/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_17/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_18/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_19/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_20/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_21/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_22/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_23/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_24/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_25/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_26/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_27/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_28/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_29/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_30/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_31/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_32/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_33/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_34/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_35/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_36/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_37/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_38/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_39/SubGenotypes.txt: No such file or directory\n", "Creating SNPMatrixAll from R Script\n", "\n", "\n", "Checking for individuals with large amounts of missing data (have >2 StDev more missing data than the average)\n", "Identifying samples with >2 stdev missing data\n", "Read 12 items\n", "Read 12 items\n", "null device \n", " 1 \n", "No samples were flagged as potentially bad samples. Would like to remove any samples from further analyses anyway? (y/n)\n", "All individuals will be analyzed.\n", "All samples were included in analyses.\n", "Reading SNPMatrixAll file\n", "Identifying monomorphic loci\n", "123 TempFiles/MonomorphicsNoParalogs.txt\n", "Updating Genotypes.txt file\n", "\n", "Identifying SNPs and determining the position of each SNP along the read.\n", "Plotting SNP locations\n", "Read 43744 items\n", "\n", "Loci have all been genotyped, and scored genotypes are stored in the file 'Genotypes.txt' in the TempFiles folder.\n", "\n", "The locations of each SNP are plotted in the file SNPLocations.pdf in the Output/RunInfo folder. Use this plot to choose a cut-off value for omitting SNPs due to artifacts toward the ends of reads.\n", "\n", "Next, run 'FilterSNPs.pl' to filter and output SNPs and monomorphic loci based on 1.) the proportion of samples genotyped at each locus and 2.) SNP positions along the reads.\n", "\n", "real\t132m18.179s\n", "user\t133m10.479s\n", "sys\t0m27.222s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl FilterSNPs.pl pctScored-17\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "maxSNP\t0\n", "MinReads\t6\n", "pctScored\t17\n", "Help\t0\n", "\n", "\n", "Running FilterSNPs.pl...\n", "\n", "Identifying all SNPs that are scored in 17% of the samples\n", "Individual SNPs, haplotypes, and monomorphic loci have been printed to files located in Output/Genotypes.\n", "These data can be further formatted using the scripts in the 'Formatting' directory\n", "\n", "real\t17m35.580s\n", "user\t17m35.781s\n", "sys\t0m0.360s\n", "\n", "Doing - lo\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl AftrRAD.pl re-TGCAG minDepth-6 P2-noP2 minIden-85 stringLength-99DataPath-Data/ BarcodePath-Barcodes/ maxProcesses-40\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "minParalog\t5\n", "minDepth\t6\n", "MaxH\t90\n", "P2\tnoP2\n", "BarcodePath\tBarcodes/\n", "minQual\t20\n", "re\tTGCAG\n", "numIndels\t3\n", "Phred\t33\n", "minIden\t85\n", "maxProcesses\t40\n", "stringLength\t99DataPath\n", "dplexedData\t0\n", "Help\t0\n", "DataPath\tData/\n", "\n", "\n", "Running AftrRAD...\n", "\n", "Recognized 1 data files.\n", "\n", "File names are...\n", " sim-lo.txt \n", "\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "The barcodes you entered for data file 0.0 (file sim-lo.txt) are...\n", " CATCATCAT TCAACTCTC AATACTAGT GAGTGACAG ATGGACCTT CGAGTTTCC TGTTAATAA TCCGTAGCC GTTTCAATC AGAATCTGC AATCGCCGT AGTCCGCTA \n", "\n", "The names you entered for data file 0.0 (file sim-lo.txt) are...\n", " 1A_0 1B_0 1C_0 1D_0 2E_0 2F_0 2G_0 2H_0 3I_0 3J_0 3K_0 3L_0 \n", "\n", "\n", "Demultiplexing samples for data file sim-lo.txt.\n", "\n", "Identifying unique sequences for each individual.\n", "23715 TempFiles/SubTemp_0/UniqueWithCountsIndividual1A_0.txt\n", "23787 TempFiles/SubTemp_0/UniqueWithCountsIndividual1B_0.txt\n", "23816 TempFiles/SubTemp_0/UniqueWithCountsIndividual1C_0.txt\n", "23743 TempFiles/SubTemp_0/UniqueWithCountsIndividual1D_0.txt\n", "23753 TempFiles/SubTemp_0/UniqueWithCountsIndividual2E_0.txt\n", "23828 TempFiles/SubTemp_0/UniqueWithCountsIndividual2F_0.txt\n", "23969 TempFiles/SubTemp_0/UniqueWithCountsIndividual2G_0.txt\n", "23582 TempFiles/SubTemp_0/UniqueWithCountsIndividual2H_0.txt\n", "23847 TempFiles/SubTemp_0/UniqueWithCountsIndividual3I_0.txt\n", "23707 TempFiles/SubTemp_0/UniqueWithCountsIndividual3J_0.txt\n", "24032 TempFiles/SubTemp_0/UniqueWithCountsIndividual3K_0.txt\n", "23685 TempFiles/SubTemp_0/UniqueWithCountsIndividual3L_0.txt\n", "\n", "Identifying all unique sequences within the dataset.\n", "\n", "Creating file to test mean read counts.\n", "Testing nonzero means in read counts to identify error reads.\n", "\n", "Performing heuristic search of all pairwise comparisons to identify potentially allelic pairs.\n", "Rapidly searching unique sequence number 1000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 2000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 3000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 4000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 5000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 6000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 7000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 8000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 9000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 10000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 11000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 12000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 13000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 14000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 15000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 16000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 17000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 18000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 19000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 20000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 21000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 22000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 23000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 24000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 25000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 26000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 27000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 28000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 29000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 30000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 31000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 32000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 33000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 34000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 35000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 36000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 37000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 38000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 39000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 40000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 41000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 42000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 43000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 44000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 45000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 46000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 47000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 48000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 49000 of 50709 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 50000 of 50709 for potentially allelic read pairs.\n", "\n", "Number of pairwise comparisons to align and evaluate is 117112\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 40000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "Working on alignment 80000 of 117112.\n", "\n", "Scoring each pairwise alignment. Alignments exceeding 85 % similarity will be retained.\n", "Parsing aligned sequences into candidate loci.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 4000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Working on alignment 8000 of 9983.\n", "Finished writing the mafft output.\n", "Identifying and removing paralogous loci.\n", "\n", "\n", "Loci have been identified. Aligned polymorphic loci are stored in the Output directory. Read counts for the alleles at these loci can be found for each individual in the files stored in TempFiles/RawReadCountFiles.\n", "\n", "Demultiplexing and other general information can be found for each dataset analyzed in the Output/RunInfo directory.\n", "\n", "Next, run Genotype.pl, followed by FilterSNPs.pl to genotype all individuals and output SNPs for subsequent analyses.\n", "\n", "\n", "real\t54m2.709s\n", "user\t58m11.067s\n", "sys\t29m20.879s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl Genotype.pl MinReads-6 subset-0 maxProcesses-40\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "subsetfile\tSamplesForSubset.txt\n", "pvalLow\t0.0001\n", "pvalThresh\t100\n", "subset\t0\n", "maxProcesses\t40\n", "MinReads\t6\n", "pvalHigh\t1e-05\n", "Help\t0\n", "\n", "\n", "Running Genotype.pl...\n", "\n", "Scoring genotypes at all nonparalogous loci.\n", "\n", "Currently genotyping sample...1C_0\t1B_0\t1D_0\t2E_0\t1A_0\t3K_0\t3I_0\t3J_0\t2G_0\t2F_0\t2H_0\t3L_0\tRunning R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "cat: TempFiles/SubGenotypes_12/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_13/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_14/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_15/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_16/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_17/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_18/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_19/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_20/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_21/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_22/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_23/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_24/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_25/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_26/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_27/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_28/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_29/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_30/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_31/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_32/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_33/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_34/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_35/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_36/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_37/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_38/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_39/SubGenotypes.txt: No such file or directory\n", "Creating SNPMatrixAll from R Script\n", "\n", "\n", "Checking for individuals with large amounts of missing data (have >2 StDev more missing data than the average)\n", "Identifying samples with >2 stdev missing data\n", "Read 12 items\n", "Read 12 items\n", "null device \n", " 1 \n", "\n", "\n", "Samples identified as potentially bad samples are \"Individual2H_0\".\n", "\n", "Do you want to remove all(a), some(s), or none(n) of these potentially bad samples from the analysis? (a/s/n)\n", "All flagged samples will be retained for analyses.\n", "Are there any additional samples you would like to remove from the analysis? (y/n)\n", "All samples were included in analyses.\n", "Reading SNPMatrixAll file\n", "Identifying monomorphic loci\n", "157 TempFiles/MonomorphicsNoParalogs.txt\n", "Updating Genotypes.txt file\n", "\n", "Identifying SNPs and determining the position of each SNP along the read.\n", "Plotting SNP locations\n", "Read 43178 items\n", "\n", "Loci have all been genotyped, and scored genotypes are stored in the file 'Genotypes.txt' in the TempFiles folder.\n", "\n", "The locations of each SNP are plotted in the file SNPLocations.pdf in the Output/RunInfo folder. Use this plot to choose a cut-off value for omitting SNPs due to artifacts toward the ends of reads.\n", "\n", "Next, run 'FilterSNPs.pl' to filter and output SNPs and monomorphic loci based on 1.) the proportion of samples genotyped at each locus and 2.) SNP positions along the reads.\n", "\n", "real\t130m1.750s\n", "user\t130m40.646s\n", "sys\t0m36.565s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl FilterSNPs.pl pctScored-17\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "maxSNP\t0\n", "MinReads\t6\n", "pctScored\t17\n", "Help\t0\n", "\n", "\n", "Running FilterSNPs.pl...\n", "\n", "Identifying all SNPs that are scored in 17% of the samples\n", "Individual SNPs, haplotypes, and monomorphic loci have been printed to files located in Output/Genotypes.\n", "These data can be further formatted using the scripts in the 'Formatting' directory\n", "\n", "real\t16m30.922s\n", "user\t16m30.948s\n", "sys\t0m0.567s\n", "\n", "Doing - hi\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl AftrRAD.pl re-TGCAG minDepth-6 P2-noP2 minIden-85 stringLength-99DataPath-Data/ BarcodePath-Barcodes/ maxProcesses-40\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "minParalog\t5\n", "minDepth\t6\n", "MaxH\t90\n", "P2\tnoP2\n", "BarcodePath\tBarcodes/\n", "minQual\t20\n", "re\tTGCAG\n", "numIndels\t3\n", "Phred\t33\n", "minIden\t85\n", "maxProcesses\t40\n", "stringLength\t99DataPath\n", "dplexedData\t0\n", "Help\t0\n", "DataPath\tData/\n", "\n", "\n", "Running AftrRAD...\n", "\n", "Recognized 1 data files.\n", "\n", "File names are...\n", " sim-hi.txt \n", "\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "The barcodes you entered for data file 0.0 (file sim-hi.txt) are...\n", " CATCATCAT TCAACTCTC AATACTAGT GAGTGACAG ATGGACCTT CGAGTTTCC TGTTAATAA TCCGTAGCC GTTTCAATC AGAATCTGC AATCGCCGT AGTCCGCTA \n", "\n", "The names you entered for data file 0.0 (file sim-hi.txt) are...\n", " 1A_0 1B_0 1C_0 1D_0 2E_0 2F_0 2G_0 2H_0 3I_0 3J_0 3K_0 3L_0 \n", "\n", "\n", "Demultiplexing samples for data file sim-hi.txt.\n", "\n", "Identifying unique sequences for each individual.\n", "23768 TempFiles/SubTemp_0/UniqueWithCountsIndividual1A_0.txt\n", "23907 TempFiles/SubTemp_0/UniqueWithCountsIndividual1B_0.txt\n", "23839 TempFiles/SubTemp_0/UniqueWithCountsIndividual1C_0.txt\n", "23943 TempFiles/SubTemp_0/UniqueWithCountsIndividual1D_0.txt\n", "23738 TempFiles/SubTemp_0/UniqueWithCountsIndividual2E_0.txt\n", "24089 TempFiles/SubTemp_0/UniqueWithCountsIndividual2F_0.txt\n", "23995 TempFiles/SubTemp_0/UniqueWithCountsIndividual2G_0.txt\n", "23937 TempFiles/SubTemp_0/UniqueWithCountsIndividual2H_0.txt\n", "23693 TempFiles/SubTemp_0/UniqueWithCountsIndividual3I_0.txt\n", "23882 TempFiles/SubTemp_0/UniqueWithCountsIndividual3J_0.txt\n", "24042 TempFiles/SubTemp_0/UniqueWithCountsIndividual3K_0.txt\n", "23819 TempFiles/SubTemp_0/UniqueWithCountsIndividual3L_0.txt\n", "\n", "Identifying all unique sequences within the dataset.\n", "\n", "Creating file to test mean read counts.\n", "Testing nonzero means in read counts to identify error reads.\n", "\n", "Performing heuristic search of all pairwise comparisons to identify potentially allelic pairs.\n", "Rapidly searching unique sequence number 1000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 2000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 3000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 4000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 5000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 6000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 7000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 8000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 9000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 10000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 11000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 12000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 13000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 14000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 15000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 16000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 17000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 18000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 19000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 20000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 21000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 22000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 23000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 24000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 25000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 26000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 27000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 28000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 29000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 30000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 31000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 32000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 33000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 34000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 35000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 36000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 37000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 38000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 39000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 40000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 41000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 42000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 43000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 44000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 45000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 46000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 47000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 48000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 49000 of 50632 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 50000 of 50632 for potentially allelic read pairs.\n", "\n", "Number of pairwise comparisons to align and evaluate is 115832\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 40000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "Working on alignment 80000 of 115832.\n", "\n", "Scoring each pairwise alignment. Alignments exceeding 85 % similarity will be retained.\n", "Parsing aligned sequences into candidate loci.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 4000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Working on alignment 8000 of 10000.\n", "Finished writing the mafft output.\n", "Identifying and removing paralogous loci.\n", "\n", "\n", "Loci have been identified. Aligned polymorphic loci are stored in the Output directory. Read counts for the alleles at these loci can be found for each individual in the files stored in TempFiles/RawReadCountFiles.\n", "\n", "Demultiplexing and other general information can be found for each dataset analyzed in the Output/RunInfo directory.\n", "\n", "Next, run Genotype.pl, followed by FilterSNPs.pl to genotype all individuals and output SNPs for subsequent analyses.\n", "\n", "\n", "real\t51m21.871s\n", "user\t55m37.453s\n", "sys\t27m33.546s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl Genotype.pl MinReads-6 subset-0 maxProcesses-40\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "subsetfile\tSamplesForSubset.txt\n", "pvalLow\t0.0001\n", "pvalThresh\t100\n", "subset\t0\n", "maxProcesses\t40\n", "MinReads\t6\n", "pvalHigh\t1e-05\n", "Help\t0\n", "\n", "\n", "Running Genotype.pl...\n", "\n", "Scoring genotypes at all nonparalogous loci.\n", "\n", "Currently genotyping sample...1B_0\t1D_0\t2F_0\t1C_0\t3L_0\t3I_0\t2H_0\t2E_0\t1A_0\t2G_0\t3K_0\t3J_0\tRunning R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "cat: TempFiles/SubGenotypes_12/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_13/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_14/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_15/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_16/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_17/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_18/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_19/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_20/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_21/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_22/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_23/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_24/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_25/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_26/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_27/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_28/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_29/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_30/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_31/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_32/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_33/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_34/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_35/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_36/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_37/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_38/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_39/SubGenotypes.txt: No such file or directory\n", "Creating SNPMatrixAll from R Script\n", "\n", "\n", "Checking for individuals with large amounts of missing data (have >2 StDev more missing data than the average)\n", "Identifying samples with >2 stdev missing data\n", "Read 12 items\n", "Read 12 items\n", "null device \n", " 1 \n", "No samples were flagged as potentially bad samples. Would like to remove any samples from further analyses anyway? (y/n)\n", "All individuals will be analyzed.\n", "All samples were included in analyses.\n", "Reading SNPMatrixAll file\n", "Identifying monomorphic loci\n", "316 TempFiles/MonomorphicsNoParalogs.txt\n", "Updating Genotypes.txt file\n", "\n", "Identifying SNPs and determining the position of each SNP along the read.\n", "Plotting SNP locations\n", "Read 42696 items\n", "\n", "Loci have all been genotyped, and scored genotypes are stored in the file 'Genotypes.txt' in the TempFiles folder.\n", "\n", "The locations of each SNP are plotted in the file SNPLocations.pdf in the Output/RunInfo folder. Use this plot to choose a cut-off value for omitting SNPs due to artifacts toward the ends of reads.\n", "\n", "Next, run 'FilterSNPs.pl' to filter and output SNPs and monomorphic loci based on 1.) the proportion of samples genotyped at each locus and 2.) SNP positions along the reads.\n", "\n", "real\t132m4.553s\n", "user\t132m49.467s\n", "sys\t0m31.332s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl FilterSNPs.pl pctScored-17\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "maxSNP\t0\n", "MinReads\t6\n", "pctScored\t17\n", "Help\t0\n", "\n", "\n", "Running FilterSNPs.pl...\n", "\n", "Identifying all SNPs that are scored in 17% of the samples\n", "Individual SNPs, haplotypes, and monomorphic loci have been printed to files located in Output/Genotypes.\n", "These data can be further formatted using the scripts in the 'Formatting' directory\n", "\n", "real\t16m55.382s\n", "user\t16m43.769s\n", "sys\t0m11.928s\n", "\n", "Doing - large\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl AftrRAD.pl re-TGCAG minDepth-6 P2-noP2 minIden-85 stringLength-99DataPath-Data/ BarcodePath-Barcodes/ maxProcesses-40\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "minParalog\t5\n", "minDepth\t6\n", "MaxH\t90\n", "P2\tnoP2\n", "BarcodePath\tBarcodes/\n", "minQual\t20\n", "re\tTGCAG\n", "numIndels\t3\n", "Phred\t33\n", "minIden\t85\n", "maxProcesses\t40\n", "stringLength\t99DataPath\n", "dplexedData\t0\n", "Help\t0\n", "DataPath\tData/\n", "\n", "\n", "Running AftrRAD...\n", "\n", "Recognized 1 data files.\n", "\n", "File names are...\n", " sim-large.txt \n", "\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "The barcodes you entered for data file 0.0 (file sim-large.txt) are...\n", " CATCATCAT TCAACTCTC AATACTAGT GAGTGACAG ATGGACCTT CGAGTTTCC TGTTAATAA TCCGTAGCC GTTTCAATC AGAATCTGC AATCGCCGT AGTCCGCTA \n", "\n", "The names you entered for data file 0.0 (file sim-large.txt) are...\n", " 1A_0 1B_0 1C_0 1D_0 2E_0 2F_0 2G_0 2H_0 3I_0 3J_0 3K_0 3L_0 \n", "\n", "\n", "Demultiplexing samples for data file sim-large.txt.\n", "\n", "Identifying unique sequences for each individual.\n", "230313 TempFiles/SubTemp_0/UniqueWithCountsIndividual1A_0.txt\n", "230281 TempFiles/SubTemp_0/UniqueWithCountsIndividual1B_0.txt\n", "230711 TempFiles/SubTemp_0/UniqueWithCountsIndividual1C_0.txt\n", "230759 TempFiles/SubTemp_0/UniqueWithCountsIndividual1D_0.txt\n", "230739 TempFiles/SubTemp_0/UniqueWithCountsIndividual2E_0.txt\n", "230332 TempFiles/SubTemp_0/UniqueWithCountsIndividual2F_0.txt\n", "230101 TempFiles/SubTemp_0/UniqueWithCountsIndividual2G_0.txt\n", "230593 TempFiles/SubTemp_0/UniqueWithCountsIndividual2H_0.txt\n", "230566 TempFiles/SubTemp_0/UniqueWithCountsIndividual3I_0.txt\n", "230382 TempFiles/SubTemp_0/UniqueWithCountsIndividual3J_0.txt\n", "230680 TempFiles/SubTemp_0/UniqueWithCountsIndividual3K_0.txt\n", "231109 TempFiles/SubTemp_0/UniqueWithCountsIndividual3L_0.txt\n", "\n", "Identifying all unique sequences within the dataset.\n", "\n", "Creating file to test mean read counts.\n", "Testing nonzero means in read counts to identify error reads.\n", "\n", "Performing heuristic search of all pairwise comparisons to identify potentially allelic pairs.\n", "Rapidly searching unique sequence number 1000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 2000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 3000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 4000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 5000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 6000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 7000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 8000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 9000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 10000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 11000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 12000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 13000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 14000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 15000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 16000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 17000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 18000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 19000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 20000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 21000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 22000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 23000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 24000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 25000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 26000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 27000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 28000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 29000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 30000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 31000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 32000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 33000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 34000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 35000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 36000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 37000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 38000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 39000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 40000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 41000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 42000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 43000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 44000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 45000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 46000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 47000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 48000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 49000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 50000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 51000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 52000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 53000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 54000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 55000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 56000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 57000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 58000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 59000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 60000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 61000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 62000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 63000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 64000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 65000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 66000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 67000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 68000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 69000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 70000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 71000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 72000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 73000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 74000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 75000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 76000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 77000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 78000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 79000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 80000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 81000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 82000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 83000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 84000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 85000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 86000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 87000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 88000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 89000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 90000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 91000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 92000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 93000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 94000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 95000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 96000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 97000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 98000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 99000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 100000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 101000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 102000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 103000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 104000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 105000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 106000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 107000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 108000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 109000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 110000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 111000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 112000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 113000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 114000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 115000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 116000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 117000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 118000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 119000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 120000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 121000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 122000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 123000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 124000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 125000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 126000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 127000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 128000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 129000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 130000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 131000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 132000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 133000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 134000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 135000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 136000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 137000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 138000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 139000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 140000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 141000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 142000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 143000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 144000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 145000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 146000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 147000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 148000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 149000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 150000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 151000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 152000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 153000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 154000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 155000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 156000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 157000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 158000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 159000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 160000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 161000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 162000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 163000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 164000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 165000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 166000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 167000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 168000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 169000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 170000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 171000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 172000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 173000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 174000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 175000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 176000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 177000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 178000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 179000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 180000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 181000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 182000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 183000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 184000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 185000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 186000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 187000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 188000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 189000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 190000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 191000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 192000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 193000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 194000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 195000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 196000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 197000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 198000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 199000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 200000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 201000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 202000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 203000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 204000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 205000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 206000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 207000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 208000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 209000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 210000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 211000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 212000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 213000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 214000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 215000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 216000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 217000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 218000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 219000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 220000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 221000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 222000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 223000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 224000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 225000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 226000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 227000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 228000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 229000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 230000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 231000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 232000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 233000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 234000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 235000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 236000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 237000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 238000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 239000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 240000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 241000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 242000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 243000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 244000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 245000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 246000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 247000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 248000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 249000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 250000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 251000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 252000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 253000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 254000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 255000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 256000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 257000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 258000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 259000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 260000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 261000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 262000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 263000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 264000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 265000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 266000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 267000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 268000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 269000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 270000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 271000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 272000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 273000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 274000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 275000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 276000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 277000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 278000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 279000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 280000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 281000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 282000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 283000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 284000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 285000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 286000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 287000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 288000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 289000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 290000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 291000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 292000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 293000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 294000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 295000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 296000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 297000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 298000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 299000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 300000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 301000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 302000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 303000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 304000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 305000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 306000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 307000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 308000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 309000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 310000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 311000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 312000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 313000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 314000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 315000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 316000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 317000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 318000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 319000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 320000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 321000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 322000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 323000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 324000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 325000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 326000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 327000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 328000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 329000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 330000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 331000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 332000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 333000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 334000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 335000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 336000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 337000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 338000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 339000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 340000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 341000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 342000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 343000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 344000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 345000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 346000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 347000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 348000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 349000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 350000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 351000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 352000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 353000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 354000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 355000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 356000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 357000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 358000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 359000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 360000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 361000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 362000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 363000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 364000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 365000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 366000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 367000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 368000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 369000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 370000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 371000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 372000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 373000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 374000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 375000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 376000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 377000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 378000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 379000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 380000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 381000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 382000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 383000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 384000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 385000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 386000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 387000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 388000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 389000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 390000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 391000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 392000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 393000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 394000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 395000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 396000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 397000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 398000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 399000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 400000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 401000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 402000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 403000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 404000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 405000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 406000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 407000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 408000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 409000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 410000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 411000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 412000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 413000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 414000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 415000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 416000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 417000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 418000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 419000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 420000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 421000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 422000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 423000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 424000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 425000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 426000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 427000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 428000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 429000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 430000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 431000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 432000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 433000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 434000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 435000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 436000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 437000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 438000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 439000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 440000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 441000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 442000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 443000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 444000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 445000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 446000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 447000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 448000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 449000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 450000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 451000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 452000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 453000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 454000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 455000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 456000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 457000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 458000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 459000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 460000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 461000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 462000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 463000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 464000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 465000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 466000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 467000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 468000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 469000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 470000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 471000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 472000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 473000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 474000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 475000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 476000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 477000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 478000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 479000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 480000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 481000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 482000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 483000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 484000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 485000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 486000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 487000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 488000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 489000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 490000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 491000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 492000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 493000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 494000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 495000 of 496426 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 496000 of 496426 for potentially allelic read pairs.\n", "\n", "Number of pairwise comparisons to align and evaluate is 1132044\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 40000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 80000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 120000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 160000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 200000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 240000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 280000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 320000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 360000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 400000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 440000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 480000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 520000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 560000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 600000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 640000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 680000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 720000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 760000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 800000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 840000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 880000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 920000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 960000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1000000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1040000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1080000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "Working on alignment 1120000 of 1132044.\n", "\n", "Scoring each pairwise alignment. Alignments exceeding 85 % similarity will be retained.\n", "Parsing aligned sequences into candidate loci.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 4000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 8000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 12000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 16000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 20000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 24000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 28000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 32000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 36000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 40000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 44000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 48000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 52000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 56000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 60000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 99096.\n", "Working on alignment 64000 of 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"Working on alignment 88000 of 99096.\n", "Working on alignment 88000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 99096.\n", "Working on alignment 92000 of 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alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Working on alignment 96000 of 99096.\n", "Finished writing the mafft output.\n", "Identifying and removing paralogous loci.\n", "\n", "\n", "Loci have been identified. Aligned polymorphic loci are stored in the Output directory. Read counts for the alleles at these loci can be found for each individual in the files stored in TempFiles/RawReadCountFiles.\n", "\n", "Demultiplexing and other general information can be found for each dataset analyzed in the Output/RunInfo directory.\n", "\n", "Next, run Genotype.pl, followed by FilterSNPs.pl to genotype all individuals and output SNPs for subsequent analyses.\n", "\n", "\n", "real\t4544m8.832s\n", "user\t4421m44.062s\n", "sys\t1322m3.702s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl Genotype.pl MinReads-6 subset-0 maxProcesses-40\n" ] } ], "source": [ "import subprocess\n", "import gzip\n", "\n", "## Set up directory structures. change the force flag if you want to\n", "## blow everything away and restart\n", "# force = True\n", "force = \"\"\n", "AFTRRAD_SIMOUT=os.path.join(AFTRRAD_DIR, \"SIMDATA/\")\n", "if force and os.path.exists(AFTRRAD_SIMOUT):\n", " shutil.rmtree(AFTRRAD_SIMOUT)\n", "if not os.path.exists(AFTRRAD_SIMOUT):\n", " os.makedirs(AFTRRAD_SIMOUT)\n", "for outdir in [\"no/\", \"lo/\", \"hi/\", \"large/\"]:\n", " tmpdir = os.path.join(AFTRRAD_SIMOUT, \"sim\"+outdir)\n", " if not os.path.exists(tmpdir):\n", " os.makedirs(tmpdir)\n", "\n", "## go to the pyrad simulated data output directory\n", "for dir in [\"no\", \"lo\", \"hi\", \"large\"]:\n", " print(\"Doing - {}\".format(dir))\n", " ## Set barcode and raw data files\n", " simdata_dir = os.path.join(SIMULATION_DATA_DIR, \"sim\"+dir)\n", " ipyrad_bcodes = os.path.join(simdata_dir, \"sim{}_barcodes.txt\".format(dir))\n", " fastqs = os.path.join(simdata_dir, \"sim{}_R1_.fastq.gz\".format(dir))\n", "\n", " simout_dir = os.path.join(AFTRRAD_SIMOUT, \"sim\"+dir)\n", " if not os.path.exists(simout_dir):\n", " os.mkdir(simout_dir)\n", " bcodes = os.path.join(simout_dir, \"aftrrad-{}-barcodes.txt\".format(dir))\n", " ## Munge the ipyrad barcodes file to the format aftrrad wants\n", " ## (barcode\\tsample name)\n", " with open(ipyrad_bcodes, 'r') as infile:\n", " with open(bcodes, 'w') as outfile:\n", " lines = infile.readlines()\n", " for line in lines:\n", " if line:\n", " outfile.write(\"\\t\".join(line.split()[::-1])+\"\\n\")\n", "\n", " ## Make data and barcodes directories\n", " data_dir = os.path.join(simout_dir, \"Data\")\n", " bcodes_dir = os.path.join(simout_dir, \"Barcodes\")\n", " for tmp in [data_dir, bcodes_dir]:\n", " if not os.path.exists(tmp):\n", " os.mkdir(tmp)\n", " ## Copy the fastq to the aftrrad/Data dir and the barcodes to the\n", " ## aftrrad/Barcodes dir. These files have to be in these directories\n", " ## and they have to have the exact same name, derp.\n", " aftrrad_fastqs = os.path.join(data_dir, \"sim-{}.txt\".format(dir))\n", " with gzip.open(fastqs, 'rb') as f_in, open(aftrrad_fastqs, 'wb') as f_out:\n", " shutil.copyfileobj(f_in, f_out)\n", " shutil.copy2(bcodes, os.path.join(bcodes_dir, \"sim-{}.txt\".format(dir)))\n", " \n", " ## AfterRAD is incredibly picky about where scripts and binaries are so you\n", " ## have to do a bunch of annoying housekeeping.\n", " cmd = \"cp -rf {}/* {}\".format(os.path.join(AFTRRAD_DIR, \"AftrRAD/AftrRADv5.0\"), simout_dir)\n", " os.system(cmd)\n", " shutil.copy2(os.path.join(AFTRRAD_DIR, \"dnaMatrix\"), simout_dir)\n", " shutil.copy2(os.path.join(AFTRRAD_DIR, \"ACANA\"), simout_dir)\n", "\n", " ## This first line of nonsense is to make it so perl can find the Parallel::ForkManager library\n", " base = \"export PATH={}perl5/bin:$PATH; cpanm --local-lib={}/perl5 local::lib && eval $(perl -I {}/perl5/lib/perl5/ -Mlocal::lib); \".format(WORK_DIR, WORK_DIR, WORK_DIR)\n", "\n", " ## Good sub for testing\n", " cmd = base + \"time perl AftrRAD.pl -h\"\n", "\n", " ## numIndels-3 What do here?\n", " ## stringLength-99 We don't care about homoplymers here, so set it very high\n", " ## maxH-90 ??\n", " ## AftrRAD manu analysis settings - P2: noP2; minDepth: 5; numIndel: 3; MinReads: 5\n", " cmd = base + \"time perl AftrRAD.pl re-TGCAG minDepth-6 P2-noP2 minIden-85 \" \\\n", " + \"stringLength-99\" \\\n", " + \"DataPath-Data/ BarcodePath-Barcodes/ maxProcesses-40\"\n", " os.chdir(simout_dir)\n", " print(cmd)\n", " output = \"\"\n", " try:\n", " output = subprocess.check_output(cmd, shell=True, stderr=subprocess.STDOUT)\n", " pass\n", " except Exception as inst:\n", " print(inst.output)\n", " print(output)\n", "\n", " ## MinReads is mindepth_statistical\n", " cmd = base + \"time perl Genotype.pl MinReads-6 subset-0 maxProcesses-40\"\n", " #cmd = base + \"perl Genotype.pl -h\"\n", " print(cmd)\n", " output = subprocess.check_output(cmd, shell=True, stderr=subprocess.STDOUT)\n", " print(output)\n", "\n", " ## pctScored is min_samples_locus as a percent of total samples, here since\n", " ## we use 2 for min_samples_locus and there are 12 simulated samples we use 17%\n", " cmd = base + \"time perl FilterSNPs.pl pctScored-17\"\n", " print(cmd)\n", " output = subprocess.check_output(cmd, shell=True, stderr=subprocess.STDOUT)\n", " print(output)" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "### Post-processing aftrrad\n", "aftrrad doesn't provide vcf as output so we have to make it ourselves\n", "\n", "Ooof. The OutputFasta.pl file has a SNPsOnly flag, but it doesn't output an alignment, so you can't use it. Also snp-sites doesn't convert ambiguity codes, so aftrRAD post processing is totally not writing out a good vcf." ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": true }, "outputs": [], "source": [ "%%bash -s \"$WORK_DIR\" -s \"$AFTRRAD_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"WORK_DIR=$1\"; export \"AFTRRAD_DIR=$2\"\n", "\n", "cd $AFTRRAD_DIR\n", "./brew/bin/brew tap homebrew/science\n", "./brew/bin/brew install snp-sites\n", "\n", "## Copy the R binary to the miniconda bin dir so it will be available\n", "cp brew/Cellar/snp-sites/2.2.0/bin/snp-sites $WORK_DIR/miniconda/bin/\n", "\n", "cd SIMDATA\n", "for d in sim*;\n", "do\n", " echo $d\n", " cd $d/Formatting\n", " perl OutputFasta.pl SNPsOnly-1 ambig-1\n", " snp-sites -v -o $d.vcf SNPMatrix_*.All.fasta\n", "done" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "### More post processing for aftrRAD\n", "The weird way snp-sites codes missing data is actually as a ref or alt\n", "allele so we have to convert all the indel variants to actually look\n", "like this './.' to align with other vcf formats for the results analysis." ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "['1', '154', '.', 'C', 'A,*', '.', '.', '.', 'GT', '0', '0', '0', '0', '1', '0', '0', '0', '0', '0', '0', './.']\n", "['1', '155', '.', 'A', 'M,*', '.', '.', '.', 'GT', '0', '1', '0', '0', '0', '0', '0', '0', '0', '0', '0', './.']\n", "['1', '156', '.', 'T', 'C,*', '.', '.', '.', 'GT', '0', '0', '0', '0', '1', '1', '1', '0', '0', '0', '0', './.']\n", "['1', '1204', '.', 'C', '*,S', '.', '.', '.', 'GT', '0', '0', '0', './.', '0', '0', '0', '0', '0', '2', '0', '0']\n", "['1', '1205', '.', 'A', '*,M', '.', '.', '.', 'GT', '0', '0', '0', './.', '0', '0', '0', '0', '2', '0', '0', '0']\n", "['1', '1206', '.', 'A', '*,T,W', '.', '.', '.', 'GT', '0', '0', '0', './.', '2', '3', '2', 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\"{}/Formatting/{}.vcf\".format(outdir, outdir)\n", " print(\"Doing - myvcf\")\n", " shutil.copy2(myvcf, myvcf+\".bak\")\n", " f = open(myvcf)\n", " lines = f.readlines()\n", " newlines = []\n", " for line in lines:\n", " if \"#\" in line:\n", " newlines.append(line.strip())\n", " continue\n", " dat = line.split()\n", " ref = dat[3]\n", " alt2 = \"\"\n", " try:\n", " alt2 = dat[4].split(\",\")[1]\n", " except:\n", " pass\n", " alt1 = dat[4].split(\",\")[0]\n", " if \"*\" in [ref, alt1, alt2]:\n", " idx = [ref, alt1, alt2].index(\"*\")\n", " for i, val in enumerate(dat[9:]):\n", " if int(val) == idx:\n", " dat[9+i] = \"./.\"\n", " #print(dat)\n", " newlines.append(\"\\t\".join(dat))\n", " outfile = open(myvcf, 'w')\n", " outfile.write(\"\\n\".join(newlines))\n" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## dDocent simulated data assembly\n", "Simulated assembly approximate runtimes:\n", "\n", "* 10000 l00bp loci ddocent runs in ~2 minutes.\n", "* 100000 100bp loci pyrad runs in ~15 minutes.\n", "\n", "~~This would be what you would do if dDocent actually could run unsupervised, but it REQUIRES\n", "you to sit there and peck at a couple keys, so this part of the script just sets up the\n", "input files and stuff for you, you're responsible for running this by hand in each sim directory.~~\n", "\n", "Actually nevermind, I found the two sections of the dDocent script that prompt for user input. These map more or less to mindepth_statistical and min_samples_locus. Hard coded two params on dDocent shell script lines 546 and 585 to set values of CUTOFF=6 and CUTOFF2=2, to align w/ these ipyrad settings.\n", "\n", "Actually nevermind, it's still doing some kind of pipe to stdout that's broken running inside the notebook, so still run these by hand. I'm leaving the hardcoded values tho.\n", "\n", "Also, on line 379 there's a hardcoded cutoff for final output where it only retains snps called in 90% of individuals.\n", " echo \"Using VCFtools to parse SNPS.vcf for SNPs that are called in at least 90% of individuals\"\n", " vcftools --vcf TotalRawSNPs.vcf --geno 0.9 --out Final --counts --recode --non-ref-af 0.001 --max-non-ref-af 0.9999 --mac 1 --minQ 30 --recode-INFO-all &>VCFtools.log\n", "\n", "Also, the final vcf files that are output contain complex genotypes rather than just snps (this is the default for freebayes), so you have to run some extra code-jams to get just\n", "the snps (vcfallelicprimitives and then vcftools --remove-indels" ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "Doing - no\n", " New Assembly: simno\n", "ipyrad -p params-simno.txt -s 1\n", "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.4.0]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: simno\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Demultiplexing fastq data to Samples\n", " [####################] 100% chunking large files | 0:00:00 \n", " [####################] 100% sorting reads | 0:00:44 \n", " [####################] 100% writing/compressing | 0:00:02 \n", "\n", "\n", "\n", "real\t0m56.558s\n", "user\t0m1.724s\n", "sys\t0m0.437s\n", "Renaming ./1A_0_R1_.fastq.gz -> Pop1_SampleA.F.fq.gz\n", "Renaming ./3J_0_R1_.fastq.gz -> Pop3_SampleJ.F.fq.gz\n", "Renaming ./1B_0_R1_.fastq.gz -> Pop1_SampleB.F.fq.gz\n", "Renaming ./3K_0_R1_.fastq.gz -> Pop3_SampleK.F.fq.gz\n", "Renaming ./2F_0_R1_.fastq.gz -> Pop2_SampleF.F.fq.gz\n", "Renaming ./1D_0_R1_.fastq.gz -> Pop1_SampleD.F.fq.gz\n", "Renaming ./1C_0_R1_.fastq.gz -> Pop1_SampleC.F.fq.gz\n", "Renaming ./3L_0_R1_.fastq.gz -> Pop3_SampleL.F.fq.gz\n", "Renaming ./2E_0_R1_.fastq.gz -> Pop2_SampleE.F.fq.gz\n", "Renaming ./3I_0_R1_.fastq.gz -> Pop3_SampleI.F.fq.gz\n", "Renaming ./2G_0_R1_.fastq.gz -> Pop2_SampleG.F.fq.gz\n", "Renaming ./2H_0_R1_.fastq.gz -> Pop2_SampleH.F.fq.gz\n", "export LD_LIBRARY_PATH=/home/iovercast/manuscript-analysis/dDocent//freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; export PATH=/home/iovercast/manuscript-analysis/dDocent/:$PATH; dDocent simno-config.txt\n", "dDocent 2.24 \n", "\n", "Contact jpuritz@gmail.com with any problems \n", "\n", " \n", "Checking for required software\n", "\n", "All required software is installed!\n", "dDocent will require input during the assembly stage. Please wait until prompt says it is safe to move program to the background.\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "Removing the _1 character and replacing with /1 in the name of every sequence\n", "Trimming reads and simultaneously assembling reference sequences\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "sort: write failed: standard output: Broken pipe\n", "sort: write error\n", "\f", " \n", " Number of Unique Sequences with More than X Coverage (Counted within individuals) \n", " \n", " 138000 +-+---------************+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " + + ***********************************+ + + + + \n", " 136000 +-+ ********* +-+ \n", " | *** | \n", " | ***** | \n", " 134000 +-+ *** +-+ \n", " | *** | \n", " 132000 +-+ * +-+ \n", " | *** | \n", " 130000 +-+ ** +-+ \n", " | ** | \n", " 128000 +-+ ** +-+ \n", " | ** | \n", " | * | \n", " 126000 +-+ *** +-+ \n", " | * | \n", " 124000 +-+ ** +-+ \n", " | * | \n", " 122000 +-+ ** +-+ \n", " | * | \n", " | **| \n", " 120000 +-+ +-+ \n", " + + + + + + + + + * \n", " 118000 +-+---------+-----------+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " 1 2 3 4 5 6 7 8 9 10 \n", " Coverage \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. In essence, you are picking a minimum (within individual) coverage level for a read (allele) to be used in the reference assembly\n", "\f", " \n", " Number of Unique Sequences present in more than X Individuals \n", " \n", " 20000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " + + + + + + + + + \n", " ****** | \n", " | *** | \n", " 18000 +-+ *** +-+ \n", " | **** | \n", " | *** | \n", " | *** | \n", " 16000 +-+ **** +-+ \n", " | * | \n", " | ******* | \n", " 14000 +-+ ***** +-+ \n", " | **** | \n", " | ***** | \n", " | **** | \n", " 12000 +-+ ** +-+ \n", " | ********* | \n", " | ***** | \n", " | ***** | \n", " 10000 +-+ ****** +-+ \n", " | ** | \n", " | ***************************| \n", " + + + + + + + + * \n", " 8000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " 2 2.5 3 3.5 4 4.5 5 5.5 6 \n", " Number of Individuals \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. Pick point right before the assymptote. A good starting cutoff might be 10% of the total number of individuals\n", "At this point, all configuration information has been entered and dDocent may take several hours to run.\n", "It is recommended that you move this script to a background operation and disable terminal input and output.\n", "All data and logfiles will still be recorded.\n", "To do this:\n", "Press control and Z simultaneously\n", "Type 'bg' without the quotes and press enter\n", "Type 'disown -h' again without the quotes and press enter\n", "\n", "Now sit back, relax, and wait for your analysis to finish.\n", "TrimmomaticSE: Started with arguments: -threads 40 -phred33 uniq.fq uniq.fq1 ILLUMINACLIP:/home/iovercast/manuscript-analysis/dDocent/TruSeq2-PE.fa:2:30:10 MINLEN:68\n", "Using PrefixPair: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT' and 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTAATGATACGGCGACCACCGAGATCTACAC'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTCAAGCAGAAGACGGCATACGA'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGCCGTATCATT'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGGTTCAGCAGGAATGCCGAGACCGATCTCGTATGCCGTCTTCTGCTTG'\n", "Using Long Clipping Sequence: 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT'\n", "ILLUMINACLIP: Using 1 prefix pairs, 6 forward/reverse sequences, 0 forward only sequences, 0 reverse only sequences\n", "Input Reads: 18850 Surviving: 18850 (100.00%) Dropped: 0 (0.00%)\n", "TrimmomaticSE: Completed successfully\n", "================================================================\n", "Program: CD-HIT, V4.6 (+OpenMP), Sep 21 2016, 20:52:31\n", "Command: cd-hit-est -i totalover.fasta -o\n", " reference.fasta.original -M 0 -T 0 -c 0.85\n", "\n", "Started: Sat Oct 1 18:01:38 2016\n", "================================================================\n", " Output \n", "----------------------------------------------------------------\n", "total number of CPUs in the system is 40\n", "Actual number of CPUs to be used: 40\n", "\n", "total seq: 18850\n", "longest and shortest : 91 and 91\n", "Total letters: 1715350\n", "Sequences have been sorted\n", "\n", "Approximated minimal memory consumption:\n", "Sequence : 4M\n", "Buffer : 40 X 12M = 491M\n", "Table : 2 X 17M = 34M\n", "Miscellaneous : 4M\n", "Total : 534M\n", "\n", "Table limit with the given memory limit:\n", "Max number of representatives: 366227\n", "Max number of word counting entries: 8747853\n", "\n", "# comparing sequences from 0 to 448\n", "---------- new table with 444 representatives\n", "# comparing sequences from 448 to 886\n", "---------- 333 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 553 to 988\n", "---------- 256 remaining sequences to the next cycle\n", "---------- new table with 171 representatives\n", "# comparing sequences from 732 to 1163\n", "---------- 189 remaining sequences to the next cycle\n", "---------- new table with 229 representatives\n", "# comparing sequences from 974 to 1399\n", "---------- 157 remaining sequences to the next cycle\n", "---------- new table with 253 representatives\n", "# comparing sequences from 1242 to 1661\n", "---------- 110 remaining sequences to the next cycle\n", "---------- new table with 282 representatives\n", "# comparing sequences from 1551 to 1962\n", "---------- 87 remaining sequences to the next cycle\n", "---------- new table with 286 representatives\n", "# comparing sequences from 1875 to 2279\n", "---------- 134 remaining sequences to the next cycle\n", "---------- new table with 239 representatives\n", "# comparing sequences from 2145 to 2542\n", "---------- 83 remaining sequences to the next cycle\n", "---------- new table with 268 representatives\n", "# comparing sequences from 2459 to 2849\n", "---------- 274 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 2575 to 2962\n", "---------- 201 remaining sequences to the next cycle\n", "---------- new table with 152 representatives\n", "# comparing sequences from 2761 to 3144\n", "---------- 260 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 2884 to 3264\n", "---------- 95 remaining sequences to the next cycle\n", "---------- new table with 232 representatives\n", "# comparing sequences from 3169 to 3542\n", "---------- 120 remaining sequences to the next cycle\n", "---------- new table with 204 representatives\n", "# comparing sequences from 3422 to 3789\n", "---------- 60 remaining sequences to the next cycle\n", "---------- new table with 256 representatives\n", "# comparing sequences from 3729 to 4089\n", "---------- 93 remaining sequences to the next cycle\n", "---------- new table with 206 representatives\n", "# comparing sequences from 3996 to 4349\n", "---------- 212 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 4137 to 4487\n", "99.8%---------- new table with 255 representatives\n", "# comparing sequences from 4487 to 4828\n", "---------- 23 remaining sequences to the next cycle\n", "---------- new table with 235 representatives\n", "# comparing sequences from 4805 to 5139\n", "---------- 111 remaining sequences to the next cycle\n", "---------- new table with 159 representatives\n", "# comparing sequences from 5028 to 5357\n", "---------- 115 remaining sequences to the next cycle\n", "---------- new table with 148 representatives\n", "# comparing sequences from 5242 to 5566\n", "---------- 165 remaining sequences to the next cycle\n", "---------- new table with 110 representatives\n", "# comparing sequences from 5401 to 5721\n", "---------- 164 remaining sequences to the next cycle\n", "---------- new table with 116 representatives\n", "# comparing sequences from 5557 to 5873\n", "---------- 141 remaining sequences to the next cycle\n", "---------- new table with 123 representatives\n", "# comparing sequences from 5732 to 6044\n", "---------- 163 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5881 to 6189\n", "---------- 141 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6048 to 6352\n", "---------- 130 remaining sequences to the next cycle\n", "---------- new table with 110 representatives\n", "# comparing sequences from 6222 to 6522\n", "---------- 79 remaining sequences to the next cycle\n", "---------- new table with 136 representatives\n", "# comparing sequences from 6443 to 6738\n", "---------- 120 remaining sequences to the next cycle\n", "---------- new table with 108 representatives\n", "# comparing sequences from 6618 to 6909\n", "---------- 107 remaining sequences to the next cycle\n", "---------- new table with 113 representatives\n", "# comparing sequences from 6802 to 7088\n", "---------- 110 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6978 to 7260\n", "---------- 107 remaining sequences to the next cycle\n", "---------- new table with 111 representatives\n", "# comparing sequences from 7153 to 7431\n", "---------- 97 remaining sequences to the next cycle\n", "---------- new table with 106 representatives\n", "# comparing sequences from 7334 to 7608\n", "---------- 75 remaining sequences to the next cycle\n", "---------- new table with 115 representatives\n", "# comparing sequences from 7533 to 7802\n", "---------- 110 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7692 to 7957\n", "---------- 86 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7871 to 8132\n", "---------- 86 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8046 to 8303\n", "---------- 66 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8237 to 8489\n", "---------- 59 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8430 to 8678\n", "---------- 34 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8644 to 8887\n", "---------- 48 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8839 to 9077\n", "---------- 34 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9043 to 9276\n", "---------- 43 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9233 to 9461\n", "---------- 39 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9422 to 9646\n", "---------- 28 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9618 to 9837\n", "---------- 16 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9821 to 10035\n", "..........---------- new table with 93 representatives\n", "# comparing sequences from 10035 to 10244\n", "........---------- new table with 100 representatives\n", "# comparing sequences from 10244 to 10448\n", "---------- 11 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 10437 to 10637\n", "...........---------- new table with 100 representatives\n", "# comparing sequences from 10637 to 10832\n", ".....---------- new table with 99 representatives\n", "# comparing sequences from 10832 to 11022\n", "........---------- new table with 77 representatives\n", "# comparing sequences from 11022 to 11208\n", ".........---------- new table with 78 representatives\n", "# comparing sequences from 11208 to 11389\n", "............---------- new table with 84 representatives\n", "# comparing sequences from 11389 to 11566\n", "...........---------- new table with 70 representatives\n", "# comparing sequences from 11566 to 11739\n", "........---------- new table with 85 representatives\n", "# comparing sequences from 11739 to 11908\n", "..........---------- new table with 71 representatives\n", "# comparing sequences from 11908 to 12073\n", "........---------- new table with 70 representatives\n", "# comparing sequences from 12073 to 12234\n", "...---------- new table with 48 representatives\n", "# comparing sequences from 12234 to 12391\n", "..........---------- new table with 70 representatives\n", "# comparing sequences from 12391 to 12544\n", ".........---------- new table with 68 representatives\n", "# comparing sequences from 12544 to 12694\n", "......---------- new table with 51 representatives\n", "# comparing sequences from 12694 to 12840\n", "...---------- new table with 46 representatives\n", "# comparing sequences from 12840 to 12983\n", "........---------- new table with 59 representatives\n", "# comparing sequences from 12983 to 13122\n", ".......---------- new table with 48 representatives\n", "# comparing sequences from 13122 to 13258\n", ".......---------- new table with 49 representatives\n", "# comparing sequences from 13258 to 13391\n", "......---------- new table with 37 representatives\n", "# comparing sequences from 13391 to 13520\n", ".......---------- new table with 41 representatives\n", "# comparing sequences from 13520 to 13646\n", ".......---------- new table with 40 representatives\n", "# comparing sequences from 13646 to 13769\n", "........---------- new table with 43 representatives\n", "# comparing sequences from 13769 to 13889\n", ".......---------- new table with 31 representatives\n", "# comparing sequences from 13889 to 14007\n", ".......---------- new table with 34 representatives\n", "# comparing sequences from 14007 to 14122\n", "...........---------- new table with 34 representatives\n", "# comparing sequences from 14122 to 14234\n", ".........---------- new table with 39 representatives\n", "# comparing sequences from 14234 to 14343\n", ".......---------- new table with 37 representatives\n", "# comparing sequences from 14343 to 14450\n", ".....---------- new table with 39 representatives\n", "# comparing sequences from 14450 to 14554\n", "......---------- new table with 35 representatives\n", "# comparing sequences from 14554 to 14656\n", ".......---------- new table with 30 representatives\n", "# comparing sequences from 14656 to 14755\n", ".......---------- new table with 29 representatives\n", "# comparing sequences from 14755 to 14852\n", ".....---------- new table with 29 representatives\n", "# comparing sequences from 14852 to 14947\n", "........---------- new table with 34 representatives\n", "# comparing sequences from 14947 to 15039\n", ".......---------- new table with 22 representatives\n", "# comparing sequences from 15039 to 15129\n", "....---------- new table with 28 representatives\n", "# comparing sequences from 15129 to 15217\n", "......---------- new table with 20 representatives\n", "# comparing sequences from 15217 to 15303\n", "....---------- new table with 30 representatives\n", "# comparing sequences from 15303 to 15387\n", ".......---------- new table with 27 representatives\n", "# comparing sequences from 15387 to 15469\n", ".......---------- new table with 23 representatives\n", "# comparing sequences from 15469 to 15549\n", "..---------- new table with 22 representatives\n", "# comparing sequences from 15549 to 15627\n", ".....---------- new table with 18 representatives\n", "# comparing sequences from 15627 to 15703\n", ".......---------- new table with 26 representatives\n", "# comparing sequences from 15703 to 15777\n", "....---------- new table with 22 representatives\n", "# comparing sequences from 15777 to 15850\n", "...---------- new table with 18 representatives\n", "# comparing sequences from 15850 to 15921\n", "......---------- new table with 19 representatives\n", "# comparing sequences from 15921 to 15990\n", "......---------- new table with 17 representatives\n", "# comparing sequences from 15990 to 16058\n", ".---------- new table with 13 representatives\n", "# comparing sequences from 16058 to 16124\n", "....---------- new table with 19 representatives\n", "# comparing sequences from 16124 to 16188\n", "..---------- new table with 18 representatives\n", "# comparing sequences from 16188 to 16251\n", "...---------- new table with 18 representatives\n", "# comparing sequences from 16251 to 16312\n", "....---------- new table with 16 representatives\n", "# comparing sequences from 16312 to 16372\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 16372 to 16431\n", "......---------- new table with 21 representatives\n", "# comparing sequences from 16431 to 16488\n", "...---------- new table with 11 representatives\n", "# comparing sequences from 16488 to 16544\n", "...---------- new table with 12 representatives\n", "# comparing sequences from 16544 to 16598\n", "..---------- new table with 10 representatives\n", "# comparing sequences from 16598 to 16651\n", "..---------- new table with 9 representatives\n", "# comparing sequences from 16651 to 16703\n", ".......---------- new table with 16 representatives\n", "# comparing sequences from 16703 to 16754\n", "....---------- new table with 15 representatives\n", "# comparing sequences from 16754 to 16803\n", ".....---------- new table with 12 representatives\n", "# comparing sequences from 16803 to 16851\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 16851 to 16898\n", ".---------- new table with 7 representatives\n", "# comparing sequences from 16898 to 16944\n", "...---------- new table with 8 representatives\n", "# comparing sequences from 16944 to 16989\n", "..---------- new table with 10 representatives\n", "# comparing sequences from 16989 to 17033\n", "...---------- new table with 10 representatives\n", "# comparing sequences from 17033 to 17076\n", "...---------- new table with 13 representatives\n", "# comparing sequences from 17076 to 17118\n", "...---------- new table with 9 representatives\n", "# comparing sequences from 17118 to 17159\n", "....---------- new table with 10 representatives\n", "# comparing sequences from 17159 to 17199\n", "....---------- new table with 12 representatives\n", "# comparing sequences from 17199 to 17238\n", "......---------- new table with 10 representatives\n", "# comparing sequences from 17238 to 17276\n", ".......---------- new table with 12 representatives\n", "# comparing sequences from 17276 to 17313\n", "...---------- new table with 8 representatives\n", "# comparing sequences from 17313 to 17349\n", "...---------- new table with 9 representatives\n", "# comparing sequences from 17349 to 17384\n", "...---------- new table with 7 representatives\n", "# comparing sequences from 17384 to 17418\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 17418 to 17452\n", ".....---------- new table with 8 representatives\n", "# comparing sequences from 17452 to 17485\n", "....---------- new table with 12 representatives\n", "# comparing sequences from 17485 to 17517\n", "..---------- new table with 5 representatives\n", "# comparing sequences from 17517 to 17548\n", "....---------- new table with 8 representatives\n", "# comparing sequences from 17548 to 17579\n", "..---------- new table with 8 representatives\n", "# comparing sequences from 17579 to 17609\n", ".....---------- new table with 9 representatives\n", "# comparing sequences from 17609 to 17638\n", "...---------- new table with 6 representatives\n", "# comparing sequences from 17638 to 17666\n", "....---------- new table with 8 representatives\n", "# comparing sequences from 17666 to 17694\n", ".....---------- new table with 8 representatives\n", "# comparing sequences from 17694 to 17721\n", "96.0%---------- new table with 3 representatives\n", "# comparing sequences from 17721 to 17747\n", "..---------- new table with 5 representatives\n", "# comparing sequences from 17747 to 17773\n", ".....---------- new table with 7 representatives\n", "# comparing sequences from 17773 to 17798\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17798 to 17823\n", ".---------- new table with 4 representatives\n", "# comparing sequences from 17823 to 17847\n", "...---------- new table with 4 representatives\n", "# comparing sequences from 17847 to 17870\n", "..---------- new table with 3 representatives\n", "# comparing sequences from 17870 to 18850\n", "...---------- new table with 186 representatives\n", "\n", " 18850 finished 10000 clusters\n", "\n", "Apprixmated maximum memory consumption: 534M\n", "writing new database\n", "writing clustering information\n", "program completed !\n", "\n", "Total CPU time 52.55\n", "[bwa_index] Pack FASTA... 0.01 sec\n", "[bwa_index] Construct BWT for the packed sequence...\n", "[bwa_index] 0.16 seconds elapse.\n", "[bwa_index] Update BWT... 0.00 sec\n", "[bwa_index] Pack forward-only FASTA... 0.02 sec\n", "[bwa_index] Construct SA from BWT and Occ... 0.08 sec\n", "[main] Version: 0.7.15-r1142-dirty\n", "[main] CMD: bwa index reference.fasta\n", "[main] Real time: 0.327 sec; CPU: 0.286 sec\n", "Using BWA to map reads.\n", "Creating alignment intervals\n", "Using FreeBayes to call SNPs\n", "Using VCFtools to parse SNPS.vcf for SNPs that are called in at least 90% of individuals\n", "Doing - lo\n", " New Assembly: simlo\n", "ipyrad -p params-simlo.txt -s 1\n", "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.4.0]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: simlo\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Demultiplexing fastq data to Samples\n", " [####################] 100% chunking large files | 0:00:00 \n", " [####################] 100% sorting reads | 0:00:45 \n", " [####################] 100% writing/compressing | 0:00:02 \n", "\n", "\n", "\n", "real\t0m58.457s\n", "user\t0m2.363s\n", "sys\t0m0.568s\n", "Renaming ./3J_0_R1_.fastq.gz -> Pop3_SampleJ.F.fq.gz\n", "Renaming ./3I_0_R1_.fastq.gz -> Pop3_SampleI.F.fq.gz\n", "Renaming ./1B_0_R1_.fastq.gz -> Pop1_SampleB.F.fq.gz\n", "Renaming ./2E_0_R1_.fastq.gz -> Pop2_SampleE.F.fq.gz\n", "Renaming ./2H_0_R1_.fastq.gz -> Pop2_SampleH.F.fq.gz\n", "Renaming ./1A_0_R1_.fastq.gz -> Pop1_SampleA.F.fq.gz\n", "Renaming ./2F_0_R1_.fastq.gz -> Pop2_SampleF.F.fq.gz\n", "Renaming ./3K_0_R1_.fastq.gz -> Pop3_SampleK.F.fq.gz\n", "Renaming ./1D_0_R1_.fastq.gz -> Pop1_SampleD.F.fq.gz\n", "Renaming ./2G_0_R1_.fastq.gz -> Pop2_SampleG.F.fq.gz\n", "Renaming ./3L_0_R1_.fastq.gz -> Pop3_SampleL.F.fq.gz\n", "Renaming ./1C_0_R1_.fastq.gz -> Pop1_SampleC.F.fq.gz\n", "export LD_LIBRARY_PATH=/home/iovercast/manuscript-analysis/dDocent//freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; export PATH=/home/iovercast/manuscript-analysis/dDocent/:$PATH; dDocent simlo-config.txt\n", "dDocent 2.24 \n", "\n", "Contact jpuritz@gmail.com with any problems \n", "\n", " \n", "Checking for required software\n", "\n", "All required software is installed!\n", "dDocent will require input during the assembly stage. Please wait until prompt says it is safe to move program to the background.\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "Removing the _1 character and replacing with /1 in the name of every sequence\n", "Trimming reads and simultaneously assembling reference sequences\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "sort: write failed: standard output: Broken pipe\n", "sort: write error\n", "\f", " \n", " Number of Unique Sequences with More than X Coverage (Counted within individuals) \n", " \n", " 138000 +-+---------************+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " + + ***********************************+ + + + + \n", " 136000 +-+ ********* +-+ \n", " | *** | \n", " | ***** | \n", " 134000 +-+ *** +-+ \n", " | *** | \n", " 132000 +-+ * +-+ \n", " | *** | \n", " 130000 +-+ ** +-+ \n", " | ** | \n", " 128000 +-+ ** +-+ \n", " | ** | \n", " | * | \n", " 126000 +-+ *** +-+ \n", " | * | \n", " 124000 +-+ ** +-+ \n", " | * | \n", " 122000 +-+ ** +-+ \n", " | * | \n", " | **| \n", " 120000 +-+ +-+ \n", " + + + + + + + + + * \n", " 118000 +-+---------+-----------+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " 1 2 3 4 5 6 7 8 9 10 \n", " Coverage \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. In essence, you are picking a minimum (within individual) coverage level for a read (allele) to be used in the reference assembly\n", "\f", " \n", " Number of Unique Sequences present in more than X Individuals \n", " \n", " 20000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " + + + + + + + + + \n", " ****** | \n", " | *** | \n", " 18000 +-+ *** +-+ \n", " | **** | \n", " | *** | \n", " | *** | \n", " 16000 +-+ **** +-+ \n", " | * | \n", " | ****** | \n", " 14000 +-+ **** +-+ \n", " | **** | \n", " | **** | \n", " | **** | \n", " 12000 +-+ **** +-+ \n", " | * | \n", " | *********** | \n", " | ****** | \n", " 10000 +-+ ******* +-+ \n", " | *** | \n", " | ***************************| \n", " + + + + + + + + * \n", " 8000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " 2 2.5 3 3.5 4 4.5 5 5.5 6 \n", " Number of Individuals \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. Pick point right before the assymptote. A good starting cutoff might be 10% of the total number of individuals\n", "At this point, all configuration information has been entered and dDocent may take several hours to run.\n", "It is recommended that you move this script to a background operation and disable terminal input and output.\n", "All data and logfiles will still be recorded.\n", "To do this:\n", "Press control and Z simultaneously\n", "Type 'bg' without the quotes and press enter\n", "Type 'disown -h' again without the quotes and press enter\n", "\n", "Now sit back, relax, and wait for your analysis to finish.\n", "TrimmomaticSE: Started with arguments: -threads 40 -phred33 uniq.fq uniq.fq1 ILLUMINACLIP:/home/iovercast/manuscript-analysis/dDocent/TruSeq2-PE.fa:2:30:10 MINLEN:68\n", "Using PrefixPair: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT' and 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTAATGATACGGCGACCACCGAGATCTACAC'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTCAAGCAGAAGACGGCATACGA'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGCCGTATCATT'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGGTTCAGCAGGAATGCCGAGACCGATCTCGTATGCCGTCTTCTGCTTG'\n", "Using Long Clipping Sequence: 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT'\n", "ILLUMINACLIP: Using 1 prefix pairs, 6 forward/reverse sequences, 0 forward only sequences, 0 reverse only sequences\n", "Input Reads: 18847 Surviving: 18847 (100.00%) Dropped: 0 (0.00%)\n", "TrimmomaticSE: Completed successfully\n", "================================================================\n", "Program: CD-HIT, V4.6 (+OpenMP), Sep 21 2016, 20:52:31\n", "Command: cd-hit-est -i totalover.fasta -o\n", " reference.fasta.original -M 0 -T 0 -c 0.85\n", "\n", "Started: Sat Oct 1 18:04:24 2016\n", "================================================================\n", " Output \n", "----------------------------------------------------------------\n", "total number of CPUs in the system is 40\n", "Actual number of CPUs to be used: 40\n", "\n", "total seq: 18847\n", "longest and shortest : 91 and 91\n", "Total letters: 1715077\n", "Sequences have been sorted\n", "\n", "Approximated minimal memory consumption:\n", "Sequence : 4M\n", "Buffer : 40 X 12M = 491M\n", "Table : 2 X 17M = 34M\n", "Miscellaneous : 4M\n", "Total : 534M\n", "\n", "Table limit with the given memory limit:\n", "Max number of representatives: 366227\n", "Max number of word counting entries: 8747853\n", "\n", "# comparing sequences from 0 to 448\n", "---------- new table with 442 representatives\n", "# comparing sequences from 448 to 886\n", "---------- 334 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 552 to 987\n", "---------- 331 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 656 to 1089\n", "---------- 327 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 762 to 1192\n", "---------- 194 remaining sequences to the next cycle\n", "---------- new table with 222 representatives\n", "# comparing sequences from 998 to 1422\n", "---------- 318 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1104 to 1526\n", "---------- 315 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1211 to 1630\n", "---------- 316 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1314 to 1731\n", "---------- 290 remaining sequences to the next cycle\n", "---------- new table with 120 representatives\n", "# comparing sequences from 1441 to 1855\n", "---------- 301 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1554 to 1965\n", "---------- 183 remaining sequences to the next cycle\n", "---------- new table with 202 representatives\n", "# comparing sequences from 1782 to 2188\n", "---------- 291 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1897 to 2300\n", "---------- 195 remaining sequences to the next cycle\n", "---------- new table with 184 representatives\n", "# comparing sequences from 2105 to 2503\n", "---------- 151 remaining sequences to the next cycle\n", "---------- new table with 212 representatives\n", "# comparing sequences from 2352 to 2744\n", "---------- 108 remaining sequences to the next cycle\n", "---------- new table with 246 representatives\n", "# comparing sequences from 2636 to 3021\n", "---------- 114 remaining sequences to the next cycle\n", "---------- new table with 219 representatives\n", "# comparing sequences from 2907 to 3286\n", "---------- 87 remaining sequences to the next cycle\n", "---------- new table with 241 representatives\n", "# comparing sequences from 3199 to 3571\n", "---------- 243 remaining sequences to the next cycle\n", "---------- new table with 105 representatives\n", "# comparing sequences from 3328 to 3697\n", "---------- 145 remaining sequences to the next cycle\n", "---------- new table with 181 representatives\n", "# comparing sequences from 3552 to 3916\n", "---------- 248 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 3668 to 4029\n", "---------- 235 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 3794 to 4152\n", "---------- 152 remaining sequences to the next cycle\n", "---------- new table with 153 representatives\n", "# comparing sequences from 4000 to 4353\n", "---------- 131 remaining sequences to the next cycle\n", "---------- new table with 164 representatives\n", "# comparing sequences from 4222 to 4570\n", "---------- 86 remaining sequences to the next cycle\n", "---------- new table with 187 representatives\n", "# comparing sequences from 4484 to 4825\n", "---------- 112 remaining sequences to the next cycle\n", "---------- new table with 164 representatives\n", "# comparing sequences from 4713 to 5049\n", "---------- 134 remaining sequences to the next cycle\n", "---------- new table with 149 representatives\n", "# comparing sequences from 4915 to 5246\n", "---------- 191 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5055 to 5383\n", "---------- 184 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5199 to 5523\n", "---------- 184 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5339 to 5660\n", "---------- 141 remaining sequences to the next cycle\n", "---------- new table with 129 representatives\n", "# comparing sequences from 5519 to 5836\n", "---------- 175 remaining sequences to the next cycle\n", "---------- new table with 103 representatives\n", "# comparing sequences from 5661 to 5974\n", "---------- 166 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5808 to 6118\n", "---------- 147 remaining sequences to the next cycle\n", "---------- new table with 103 representatives\n", "# comparing sequences from 5971 to 6277\n", "---------- 143 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6134 to 6436\n", "---------- 143 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6293 to 6591\n", "---------- 135 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6456 to 6751\n", "---------- 134 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6617 to 6908\n", "---------- 128 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6780 to 7067\n", "---------- 126 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6941 to 7224\n", "---------- 115 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7109 to 7388\n", "---------- 120 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7268 to 7543\n", "---------- 106 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7437 to 7708\n", "---------- 100 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7608 to 7875\n", "---------- 99 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7776 to 8039\n", "---------- 76 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7963 to 8222\n", "---------- 87 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8135 to 8390\n", "---------- 50 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8340 to 8590\n", "---------- 59 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8531 to 8776\n", "---------- 58 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8718 to 8959\n", "---------- 32 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8927 to 9163\n", "---------- 28 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9135 to 9366\n", "---------- 45 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9321 to 9547\n", "---------- 36 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9511 to 9733\n", "---------- 28 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9705 to 9922\n", "---------- 6 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9916 to 10128\n", "---------- 4 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 10124 to 10331\n", ".......---------- new table with 99 representatives\n", "# comparing sequences from 10331 to 10533\n", "---------- 6 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 10527 to 10725\n", "...........---------- new table with 97 representatives\n", "# comparing sequences from 10725 to 10918\n", ".........---------- new table with 86 representatives\n", "# comparing sequences from 10918 to 11106\n", "......---------- new table with 70 representatives\n", "# comparing sequences from 11106 to 11290\n", "............---------- new table with 82 representatives\n", "# comparing sequences from 11290 to 11469\n", "........---------- new table with 74 representatives\n", "# comparing sequences from 11469 to 11644\n", "......---------- new table with 76 representatives\n", "# comparing sequences from 11644 to 11815\n", ".........---------- new table with 75 representatives\n", "# comparing sequences from 11815 to 11982\n", "........---------- new table with 77 representatives\n", "# comparing sequences from 11982 to 12145\n", ".....---------- new table with 56 representatives\n", "# comparing sequences from 12145 to 12304\n", "......---------- new table with 58 representatives\n", "# comparing sequences from 12304 to 12459\n", ".............---------- new table with 69 representatives\n", "# comparing sequences from 12459 to 12611\n", "....---------- new table with 55 representatives\n", "# comparing sequences from 12611 to 12759\n", "......---------- new table with 49 representatives\n", "# comparing sequences from 12759 to 12903\n", ".....---------- new table with 49 representatives\n", "# comparing sequences from 12903 to 13044\n", "..........---------- new table with 54 representatives\n", "# comparing sequences from 13044 to 13182\n", "..........---------- new table with 52 representatives\n", "# comparing sequences from 13182 to 13316\n", ".....---------- new table with 39 representatives\n", "# comparing sequences from 13316 to 13447\n", "....---------- new table with 38 representatives\n", "# comparing sequences from 13447 to 13575\n", ".....---------- new table with 47 representatives\n", "# comparing sequences from 13575 to 13700\n", ".....---------- new table with 41 representatives\n", "# comparing sequences from 13700 to 13822\n", ".......---------- new table with 35 representatives\n", "# comparing sequences from 13822 to 13941\n", ".....---------- new table with 30 representatives\n", "# comparing sequences from 13941 to 14057\n", "............---------- new table with 42 representatives\n", "# comparing sequences from 14057 to 14171\n", ".........---------- new table with 35 representatives\n", "# comparing sequences from 14171 to 14282\n", "......---------- new table with 36 representatives\n", "# comparing sequences from 14282 to 14390\n", ".........---------- new table with 38 representatives\n", "# comparing sequences from 14390 to 14496\n", ".....---------- new table with 36 representatives\n", "# comparing sequences from 14496 to 14599\n", ".......---------- new table with 34 representatives\n", "# comparing sequences from 14599 to 14700\n", "........---------- new table with 27 representatives\n", "# comparing sequences from 14700 to 14798\n", "....---------- new table with 33 representatives\n", "# comparing sequences from 14798 to 14894\n", ".......---------- new table with 31 representatives\n", "# comparing sequences from 14894 to 14988\n", ".......---------- new table with 28 representatives\n", "# comparing sequences from 14988 to 15079\n", ".......---------- new table with 28 representatives\n", "# comparing sequences from 15079 to 15168\n", "........---------- new table with 22 representatives\n", "# comparing sequences from 15168 to 15255\n", ".......---------- new table with 25 representatives\n", "# comparing sequences from 15255 to 15340\n", "..........---------- new table with 30 representatives\n", "# comparing sequences from 15340 to 15423\n", ".......---------- new table with 27 representatives\n", "# comparing sequences from 15423 to 15504\n", "......---------- new table with 22 representatives\n", "# comparing sequences from 15504 to 15583\n", "........---------- new table with 23 representatives\n", "# comparing sequences from 15583 to 15660\n", ".....---------- new table with 21 representatives\n", "# comparing sequences from 15660 to 15735\n", "........---------- new table with 20 representatives\n", "# comparing sequences from 15735 to 15809\n", ".......---------- new table with 22 representatives\n", "# comparing sequences from 15809 to 15881\n", "....---------- new table with 18 representatives\n", "# comparing sequences from 15881 to 15951\n", "......---------- new table with 19 representatives\n", "# comparing sequences from 15951 to 16019\n", "..---------- new table with 13 representatives\n", "# comparing sequences from 16019 to 16086\n", ".....---------- new table with 15 representatives\n", "# comparing sequences from 16086 to 16151\n", ".......---------- new table with 17 representatives\n", "# comparing sequences from 16151 to 16215\n", "....---------- new table with 19 representatives\n", "# comparing sequences from 16215 to 16277\n", "......---------- new table with 15 representatives\n", "# comparing sequences from 16277 to 16338\n", "....---------- new table with 15 representatives\n", "# comparing sequences from 16338 to 16397\n", "...---------- new table with 17 representatives\n", "# comparing sequences from 16397 to 16455\n", "..---------- new table with 17 representatives\n", "# comparing sequences from 16455 to 16511\n", ".....---------- new table with 11 representatives\n", "# comparing sequences from 16511 to 16566\n", "99.9%---------- new table with 6 representatives\n", "# comparing sequences from 16566 to 16620\n", ".........---------- new table with 14 representatives\n", "# comparing sequences from 16620 to 16673\n", ".....---------- new table with 12 representatives\n", "# comparing sequences from 16673 to 16724\n", ".....---------- new table with 16 representatives\n", "# comparing sequences from 16724 to 16774\n", "...---------- new table with 13 representatives\n", "# comparing sequences from 16774 to 16823\n", "....---------- new table with 11 representatives\n", "# comparing sequences from 16823 to 16871\n", "....---------- new table with 11 representatives\n", "# comparing sequences from 16871 to 16918\n", "......---------- new table with 11 representatives\n", "# comparing sequences from 16918 to 16963\n", "..---------- new table with 8 representatives\n", "# comparing sequences from 16963 to 17007\n", "...---------- new table with 10 representatives\n", "# comparing sequences from 17007 to 17050\n", "....---------- new table with 9 representatives\n", "# comparing sequences from 17050 to 17092\n", "...---------- new table with 14 representatives\n", "# comparing sequences from 17092 to 17133\n", "...---------- new table with 6 representatives\n", "# comparing sequences from 17133 to 17173\n", "......---------- new table with 13 representatives\n", "# comparing sequences from 17173 to 17212\n", ".....---------- new table with 12 representatives\n", "# comparing sequences from 17212 to 17250\n", ".....---------- new table with 9 representatives\n", "# comparing sequences from 17250 to 17288\n", ".......---------- new table with 10 representatives\n", "# comparing sequences from 17288 to 17325\n", ".......---------- new table with 11 representatives\n", "# comparing sequences from 17325 to 17361\n", "....---------- new table with 8 representatives\n", "# comparing sequences from 17361 to 17396\n", "......---------- new table with 11 representatives\n", "# comparing sequences from 17396 to 17430\n", "...---------- new table with 9 representatives\n", "# comparing sequences from 17430 to 17463\n", "...---------- new table with 11 representatives\n", "# comparing sequences from 17463 to 17495\n", "....---------- new table with 7 representatives\n", "# comparing sequences from 17495 to 17527\n", "....---------- new table with 6 representatives\n", "# comparing sequences from 17527 to 17558\n", "...---------- new table with 7 representatives\n", "# comparing sequences from 17558 to 17588\n", "......---------- new table with 11 representatives\n", "# comparing sequences from 17588 to 17617\n", "..---------- new table with 5 representatives\n", "# comparing sequences from 17617 to 17646\n", ".....---------- new table with 8 representatives\n", "# comparing sequences from 17646 to 17674\n", "......---------- new table with 10 representatives\n", "# comparing sequences from 17674 to 17701\n", ".---------- new table with 2 representatives\n", "# comparing sequences from 17701 to 17728\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17728 to 17754\n", ".....---------- new table with 7 representatives\n", "# comparing sequences from 17754 to 17780\n", "....---------- new table with 7 representatives\n", "# comparing sequences from 17780 to 17805\n", "...---------- new table with 3 representatives\n", "# comparing sequences from 17805 to 17829\n", ".---------- new table with 5 representatives\n", "# comparing sequences from 17829 to 17853\n", ".---------- new table with 2 representatives\n", "# comparing sequences from 17853 to 18847\n", "....---------- new table with 190 representatives\n", "\n", " 18847 finished 10000 clusters\n", "\n", "Apprixmated maximum memory consumption: 534M\n", "writing new database\n", "writing clustering information\n", "program completed !\n", "\n", "Total CPU time 55.78\n", "[bwa_index] Pack FASTA... 0.01 sec\n", "[bwa_index] Construct BWT for the packed sequence...\n", "[bwa_index] 0.16 seconds elapse.\n", "[bwa_index] Update BWT... 0.01 sec\n", "[bwa_index] Pack forward-only FASTA... 0.01 sec\n", "[bwa_index] Construct SA from BWT and Occ... 0.09 sec\n", "[main] Version: 0.7.15-r1142-dirty\n", "[main] CMD: bwa index reference.fasta\n", "[main] Real time: 0.338 sec; CPU: 0.299 sec\n", "Using BWA to map reads.\n", "Creating alignment intervals\n", "Using FreeBayes to call SNPs\n", "Using VCFtools to parse SNPS.vcf for SNPs that are called in at least 90% of individuals\n", "Doing - hi\n", " New Assembly: simhi\n", "ipyrad -p params-simhi.txt -s 1\n", "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.4.0]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: simhi\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Demultiplexing fastq data to Samples\n", " [####################] 100% chunking large files | 0:00:00 \n", " [####################] 100% sorting reads | 0:00:45 \n", " [####################] 100% writing/compressing | 0:00:02 \n", "\n", "\n", "\n", "real\t0m57.199s\n", "user\t0m1.775s\n", "sys\t0m0.522s\n", "Renaming ./3K_0_R1_.fastq.gz -> Pop3_SampleK.F.fq.gz\n", "Renaming ./2G_0_R1_.fastq.gz -> Pop2_SampleG.F.fq.gz\n", "Renaming ./2E_0_R1_.fastq.gz -> Pop2_SampleE.F.fq.gz\n", "Renaming ./3L_0_R1_.fastq.gz -> Pop3_SampleL.F.fq.gz\n", "Renaming ./1A_0_R1_.fastq.gz -> Pop1_SampleA.F.fq.gz\n", "Renaming ./2H_0_R1_.fastq.gz -> Pop2_SampleH.F.fq.gz\n", "Renaming ./3J_0_R1_.fastq.gz -> Pop3_SampleJ.F.fq.gz\n", "Renaming ./1B_0_R1_.fastq.gz -> Pop1_SampleB.F.fq.gz\n", "Renaming ./1D_0_R1_.fastq.gz -> Pop1_SampleD.F.fq.gz\n", "Renaming ./2F_0_R1_.fastq.gz -> Pop2_SampleF.F.fq.gz\n", "Renaming ./3I_0_R1_.fastq.gz -> Pop3_SampleI.F.fq.gz\n", "Renaming ./1C_0_R1_.fastq.gz -> Pop1_SampleC.F.fq.gz\n", "export LD_LIBRARY_PATH=/home/iovercast/manuscript-analysis/dDocent//freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; export PATH=/home/iovercast/manuscript-analysis/dDocent/:$PATH; dDocent simhi-config.txt\n", "dDocent 2.24 \n", "\n", "Contact jpuritz@gmail.com with any problems \n", "\n", " \n", "Checking for required software\n", "\n", "All required software is installed!\n", "dDocent will require input during the assembly stage. Please wait until prompt says it is safe to move program to the background.\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "Removing the _1 character and replacing with /1 in the name of every sequence\n", "Trimming reads and simultaneously assembling reference sequences\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "sort: write failed: standard output: Broken pipe\n", "sort: write error\n", "\f", " \n", " Number of Unique Sequences with More than X Coverage (Counted within individuals) \n", " \n", " 138000 +-+---------************+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " + + ***********************************+ + + + + \n", " 136000 +-+ ********* +-+ \n", " | *** | \n", " | ***** | \n", " 134000 +-+ *** +-+ \n", " | *** | \n", " 132000 +-+ * +-+ \n", " | *** | \n", " 130000 +-+ ** +-+ \n", " | ** | \n", " 128000 +-+ ** +-+ \n", " | ** | \n", " | * | \n", " 126000 +-+ *** +-+ \n", " | * | \n", " 124000 +-+ ** +-+ \n", " | * | \n", " 122000 +-+ ** +-+ \n", " | * | \n", " | **| \n", " 120000 +-+ +-+ \n", " + + + + + + + + + * \n", " 118000 +-+---------+-----------+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " 1 2 3 4 5 6 7 8 9 10 \n", " Coverage \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. In essence, you are picking a minimum (within individual) coverage level for a read (allele) to be used in the reference assembly\n", "\f", " \n", " Number of Unique Sequences present in more than X Individuals \n", " \n", " 20000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " + + + + + + + + + \n", " ****** | \n", " | *** | \n", " 18000 +-+ *** +-+ \n", " | **** | \n", " | *** | \n", " | *** | \n", " 16000 +-+ **** +-+ \n", " | * | \n", " | ******* | \n", " 14000 +-+ ***** +-+ \n", " | **** | \n", " | ***** | \n", " | **** | \n", " 12000 +-+ ** +-+ \n", " | ********* | \n", " | ***** | \n", " | ***** | \n", " 10000 +-+ ****** +-+ \n", " | ** | \n", " | ***************************| \n", " + + + + + + + + * \n", " 8000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " 2 2.5 3 3.5 4 4.5 5 5.5 6 \n", " Number of Individuals \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. Pick point right before the assymptote. A good starting cutoff might be 10% of the total number of individuals\n", "At this point, all configuration information has been entered and dDocent may take several hours to run.\n", "It is recommended that you move this script to a background operation and disable terminal input and output.\n", "All data and logfiles will still be recorded.\n", "To do this:\n", "Press control and Z simultaneously\n", "Type 'bg' without the quotes and press enter\n", "Type 'disown -h' again without the quotes and press enter\n", "\n", "Now sit back, relax, and wait for your analysis to finish.\n", "TrimmomaticSE: Started with arguments: -threads 40 -phred33 uniq.fq uniq.fq1 ILLUMINACLIP:/home/iovercast/manuscript-analysis/dDocent/TruSeq2-PE.fa:2:30:10 MINLEN:68\n", "Using PrefixPair: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT' and 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTAATGATACGGCGACCACCGAGATCTACAC'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTCAAGCAGAAGACGGCATACGA'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGCCGTATCATT'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGGTTCAGCAGGAATGCCGAGACCGATCTCGTATGCCGTCTTCTGCTTG'\n", "Using Long Clipping Sequence: 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT'\n", "ILLUMINACLIP: Using 1 prefix pairs, 6 forward/reverse sequences, 0 forward only sequences, 0 reverse only sequences\n", "Input Reads: 18867 Surviving: 18867 (100.00%) Dropped: 0 (0.00%)\n", "TrimmomaticSE: Completed successfully\n", "================================================================\n", "Program: CD-HIT, V4.6 (+OpenMP), Sep 21 2016, 20:52:31\n", "Command: cd-hit-est -i totalover.fasta -o\n", " reference.fasta.original -M 0 -T 0 -c 0.85\n", "\n", "Started: Sat Oct 1 18:07:19 2016\n", "================================================================\n", " Output \n", "----------------------------------------------------------------\n", "total number of CPUs in the system is 40\n", "Actual number of CPUs to be used: 40\n", "\n", "total seq: 18867\n", "longest and shortest : 91 and 91\n", "Total letters: 1716897\n", "Sequences have been sorted\n", "\n", "Approximated minimal memory consumption:\n", "Sequence : 4M\n", "Buffer : 40 X 12M = 491M\n", "Table : 2 X 17M = 34M\n", "Miscellaneous : 4M\n", "Total : 534M\n", "\n", "Table limit with the given memory limit:\n", "Max number of representatives: 366227\n", "Max number of word counting entries: 8747853\n", "\n", "# comparing sequences from 0 to 449\n", "---------- new table with 444 representatives\n", "# comparing sequences from 449 to 887\n", "---------- 334 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 553 to 989\n", "---------- 241 remaining sequences to the next cycle\n", "---------- new table with 185 representatives\n", "# comparing sequences from 748 to 1179\n", "---------- 239 remaining sequences to the next cycle\n", "---------- new table with 181 representatives\n", "# comparing sequences from 940 to 1366\n", "---------- 206 remaining sequences to the next cycle\n", "---------- new table with 205 representatives\n", "# comparing sequences from 1160 to 1581\n", "---------- 205 remaining sequences to the next cycle\n", "---------- new table with 202 representatives\n", "# comparing sequences from 1376 to 1792\n", "---------- 200 remaining sequences to the next cycle\n", "---------- new table with 197 representatives\n", "# comparing sequences from 1592 to 2003\n", "---------- 195 remaining sequences to the next cycle\n", "---------- new table with 196 representatives\n", "# comparing sequences from 1808 to 2214\n", "---------- 172 remaining sequences to the next cycle\n", "---------- new table with 204 representatives\n", "# comparing sequences from 2042 to 2442\n", "---------- 215 remaining sequences to the next cycle\n", "---------- new table with 160 representatives\n", "# comparing sequences from 2227 to 2623\n", "---------- 188 remaining sequences to the next cycle\n", "---------- new table with 177 representatives\n", "# comparing sequences from 2435 to 2826\n", "---------- 204 remaining sequences to the next cycle\n", "---------- new table with 162 representatives\n", "# comparing sequences from 2622 to 3008\n", "---------- 177 remaining sequences to the next cycle\n", "---------- new table with 171 representatives\n", "# comparing sequences from 2831 to 3212\n", "---------- 91 remaining sequences to the next cycle\n", "---------- new table with 235 representatives\n", "# comparing sequences from 3121 to 3495\n", "---------- 122 remaining sequences to the next cycle\n", "---------- new table with 207 representatives\n", "# comparing sequences from 3373 to 3741\n", "---------- 248 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 3493 to 3859\n", "---------- 113 remaining sequences to the next cycle\n", "---------- new table with 211 representatives\n", "# comparing sequences from 3746 to 4106\n", "---------- 115 remaining sequences to the next cycle\n", "---------- new table with 187 representatives\n", "# comparing sequences from 3991 to 4345\n", "80.8%---------- new table with 258 representatives\n", "# comparing sequences from 4345 to 4690\n", "---------- 127 remaining sequences to the next cycle\n", "---------- new table with 158 representatives\n", "# comparing sequences from 4563 to 4903\n", "---------- 154 remaining sequences to the next cycle\n", "---------- new table with 142 representatives\n", "# comparing sequences from 4749 to 5085\n", "---------- 199 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 4886 to 5218\n", "---------- 189 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5029 to 5358\n", "---------- 183 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5175 to 5501\n", "100.0%---------- new table with 230 representatives\n", "# comparing sequences from 5501 to 5819\n", "86.1%---------- new table with 219 representatives\n", "# comparing sequences from 5819 to 6129\n", "---------- 39 remaining sequences to the next cycle\n", "---------- new table with 166 representatives\n", "# comparing sequences from 6090 to 6394\n", "---------- 52 remaining sequences to the next cycle\n", "---------- new table with 155 representatives\n", "# comparing sequences from 6342 to 6640\n", "---------- 140 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6500 to 6794\n", "---------- 77 remaining sequences to the next cycle\n", "---------- new table with 131 representatives\n", "# comparing sequences from 6717 to 7006\n", "---------- 41 remaining sequences to the next cycle\n", "---------- new table with 145 representatives\n", "# comparing sequences from 6965 to 7248\n", "---------- 64 remaining sequences to the next cycle\n", "---------- new table with 142 representatives\n", "# comparing sequences from 7184 to 7462\n", "---------- 52 remaining sequences to the next cycle\n", "---------- new table with 132 representatives\n", "# comparing sequences from 7410 to 7682\n", "---------- 96 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7586 to 7854\n", "---------- 97 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7757 to 8021\n", "---------- 93 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7928 to 8188\n", "---------- 90 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8098 to 8354\n", "---------- 52 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8302 to 8553\n", "---------- 77 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8476 to 8723\n", "---------- 44 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8679 to 8921\n", "---------- 33 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8888 to 9125\n", "---------- 50 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9075 to 9308\n", "---------- 51 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9257 to 9485\n", "99.9%---------- new table with 115 representatives\n", "# comparing sequences from 9485 to 9708\n", "---------- 21 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9687 to 9905\n", "---------- 11 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9894 to 10107\n", "........---------- new table with 97 representatives\n", "# comparing sequences from 10107 to 10315\n", "...........---------- new table with 94 representatives\n", "# comparing sequences from 10315 to 10518\n", "---------- 9 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 10509 to 10708\n", ".............---------- new table with 94 representatives\n", "# comparing sequences from 10708 to 10902\n", ".............---------- new table with 92 representatives\n", "# comparing sequences from 10902 to 11091\n", ".......---------- new table with 73 representatives\n", "# comparing sequences from 11091 to 11276\n", "..........---------- new table with 84 representatives\n", "# comparing sequences from 11276 to 11456\n", "..........---------- new table with 79 representatives\n", "# comparing sequences from 11456 to 11632\n", "....---------- new table with 62 representatives\n", "# comparing sequences from 11632 to 11804\n", ".............---------- new table with 85 representatives\n", "# comparing sequences from 11804 to 11972\n", ".........---------- new table with 69 representatives\n", "# comparing sequences from 11972 to 12136\n", "....---------- new table with 61 representatives\n", "# comparing sequences from 12136 to 12296\n", ".....---------- new table with 55 representatives\n", "# comparing sequences from 12296 to 12452\n", "...........---------- new table with 69 representatives\n", "# comparing sequences from 12452 to 12604\n", ".......---------- new table with 61 representatives\n", "# comparing sequences from 12604 to 12753\n", ".......---------- new table with 50 representatives\n", "# comparing sequences from 12753 to 12898\n", "...---------- new table with 50 representatives\n", "# comparing sequences from 12898 to 13040\n", "......---------- new table with 58 representatives\n", "# comparing sequences from 13040 to 13178\n", "........---------- new table with 53 representatives\n", "# comparing sequences from 13178 to 13313\n", "......---------- new table with 39 representatives\n", "# comparing sequences from 13313 to 13445\n", "........---------- new table with 42 representatives\n", "# comparing sequences from 13445 to 13574\n", ".....---------- new table with 40 representatives\n", "# comparing sequences from 13574 to 13700\n", ".......---------- new table with 43 representatives\n", "# comparing sequences from 13700 to 13823\n", ".....---------- new table with 37 representatives\n", "# comparing sequences from 13823 to 13943\n", "....---------- new table with 33 representatives\n", "# comparing sequences from 13943 to 14060\n", ".......---------- new table with 37 representatives\n", "# comparing sequences from 14060 to 14174\n", "......---------- new table with 36 representatives\n", "# comparing sequences from 14174 to 14285\n", ".....---------- new table with 42 representatives\n", "# comparing sequences from 14285 to 14394\n", ".....---------- new table with 37 representatives\n", "# comparing sequences from 14394 to 14500\n", "....---------- new table with 34 representatives\n", "# comparing sequences from 14500 to 14603\n", "....---------- new table with 30 representatives\n", "# comparing sequences from 14603 to 14704\n", "........---------- new table with 33 representatives\n", "# comparing sequences from 14704 to 14803\n", "..........---------- new table with 34 representatives\n", "# comparing sequences from 14803 to 14899\n", "......---------- new table with 30 representatives\n", "# comparing sequences from 14899 to 14993\n", ".......---------- new table with 30 representatives\n", "# comparing sequences from 14993 to 15085\n", "....---------- new table with 26 representatives\n", "# comparing sequences from 15085 to 15175\n", ".....---------- new table with 25 representatives\n", "# comparing sequences from 15175 to 15262\n", ".....---------- new table with 24 representatives\n", "# comparing sequences from 15262 to 15347\n", "...---------- new table with 27 representatives\n", "# comparing sequences from 15347 to 15430\n", ".....---------- new table with 25 representatives\n", "# comparing sequences from 15430 to 15511\n", "...---------- new table with 22 representatives\n", "# comparing sequences from 15511 to 15590\n", ".......---------- new table with 22 representatives\n", "# comparing sequences from 15590 to 15668\n", "......---------- new table with 22 representatives\n", "# comparing sequences from 15668 to 15744\n", "......---------- new table with 20 representatives\n", "# comparing sequences from 15744 to 15818\n", "........---------- new table with 25 representatives\n", "# comparing sequences from 15818 to 15890\n", "....---------- new table with 12 representatives\n", "# comparing sequences from 15890 to 15960\n", "....---------- new table with 17 representatives\n", "# comparing sequences from 15960 to 16029\n", ".....---------- new table with 16 representatives\n", "# comparing sequences from 16029 to 16096\n", ".---------- new table with 16 representatives\n", "# comparing sequences from 16096 to 16161\n", "...---------- new table with 17 representatives\n", "# comparing sequences from 16161 to 16225\n", "......---------- new table with 18 representatives\n", "# comparing sequences from 16225 to 16287\n", "....---------- new table with 17 representatives\n", "# comparing sequences from 16287 to 16348\n", ".---------- new table with 12 representatives\n", "# comparing sequences from 16348 to 16407\n", "....---------- new table with 16 representatives\n", "# comparing sequences from 16407 to 16465\n", "...---------- new table with 14 representatives\n", "# comparing sequences from 16465 to 16522\n", ".......---------- new table with 14 representatives\n", "# comparing sequences from 16522 to 16577\n", "...---------- new table with 10 representatives\n", "# comparing sequences from 16577 to 16631\n", ".---------- new table with 14 representatives\n", "# comparing sequences from 16631 to 16684\n", "...---------- new table with 10 representatives\n", "# comparing sequences from 16684 to 16735\n", ".....---------- new table with 15 representatives\n", "# comparing sequences from 16735 to 16785\n", ".....---------- new table with 14 representatives\n", "# comparing sequences from 16785 to 16834\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 16834 to 16882\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 16882 to 16929\n", ".---------- new table with 6 representatives\n", "# comparing sequences from 16929 to 16975\n", ".......---------- new table with 13 representatives\n", "# comparing sequences from 16975 to 17020\n", "....---------- new table with 9 representatives\n", "# comparing sequences from 17020 to 17063\n", "..---------- new table with 10 representatives\n", "# comparing sequences from 17063 to 17105\n", "......---------- new table with 11 representatives\n", "# comparing sequences from 17105 to 17146\n", "..---------- new table with 7 representatives\n", "# comparing sequences from 17146 to 17186\n", "...---------- new table with 12 representatives\n", "# comparing sequences from 17186 to 17226\n", ".---------- new table with 9 representatives\n", "# comparing sequences from 17226 to 17265\n", ".....---------- new table with 10 representatives\n", "# comparing sequences from 17265 to 17303\n", ".........---------- new table with 12 representatives\n", "# comparing sequences from 17303 to 17340\n", "....---------- new table with 9 representatives\n", "# comparing sequences from 17340 to 17376\n", ".....---------- new table with 8 representatives\n", "# comparing sequences from 17376 to 17411\n", ".....---------- new table with 9 representatives\n", "# comparing sequences from 17411 to 17445\n", "......---------- new table with 12 representatives\n", "# comparing sequences from 17445 to 17478\n", "...---------- new table with 11 representatives\n", "# comparing sequences from 17478 to 17511\n", ".........---------- new table with 12 representatives\n", "# comparing sequences from 17511 to 17543\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17543 to 17574\n", "...---------- new table with 8 representatives\n", "# comparing sequences from 17574 to 17604\n", "...---------- new table with 9 representatives\n", "# comparing sequences from 17604 to 17634\n", "...---------- new table with 6 representatives\n", "# comparing sequences from 17634 to 17663\n", ".---------- new table with 4 representatives\n", "# comparing sequences from 17663 to 17691\n", "....---------- new table with 11 representatives\n", "# comparing sequences from 17691 to 17719\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17719 to 17746\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17746 to 17772\n", "....---------- new table with 5 representatives\n", "# comparing sequences from 17772 to 17798\n", "...---------- new table with 7 representatives\n", "# comparing sequences from 17798 to 17823\n", "---------- new table with 3 representatives\n", "# comparing sequences from 17823 to 17847\n", "....---------- new table with 6 representatives\n", "# comparing sequences from 17847 to 17871\n", "95.8%---------- new table with 1 representatives\n", "# comparing sequences from 17871 to 18867\n", ".....---------- new table with 189 representatives\n", "\n", " 18867 finished 10000 clusters\n", "\n", "Apprixmated maximum memory consumption: 534M\n", "writing new database\n", "writing clustering information\n", "program completed !\n", "\n", "Total CPU time 53.40\n", "[bwa_index] Pack FASTA... 0.01 sec\n", "[bwa_index] Construct BWT for the packed sequence...\n", "[bwa_index] 0.24 seconds elapse.\n", "[bwa_index] Update BWT... 0.01 sec\n", "[bwa_index] Pack forward-only FASTA... 0.02 sec\n", "[bwa_index] Construct SA from BWT and Occ... 0.08 sec\n", "[main] Version: 0.7.15-r1142-dirty\n", "[main] CMD: bwa index reference.fasta\n", "[main] Real time: 0.409 sec; CPU: 0.369 sec\n", "Using BWA to map reads.\n", "Creating alignment intervals\n", "Using FreeBayes to call SNPs\n", "Using VCFtools to parse SNPS.vcf for SNPs that are called in at least 90% of individuals\n", "Doing - large\n", " New Assembly: simlarge\n", "ipyrad -p params-simlarge.txt -s 1\n", "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.4.0]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: simlarge\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Demultiplexing fastq data to Samples\n", " [####################] 100% chunking large files | 0:01:06 \n", " [####################] 100% sorting reads | 0:00:59 \n", " [####################] 100% writing/compressing | 0:00:11 \n", "\n", "\n", "\n", "real\t2m28.462s\n", "user\t0m2.154s\n", "sys\t0m1.422s\n", "Renaming ./3K_0_R1_.fastq.gz -> Pop3_SampleK.F.fq.gz\n", "Renaming ./2H_0_R1_.fastq.gz -> Pop2_SampleH.F.fq.gz\n", "Renaming ./1B_0_R1_.fastq.gz -> Pop1_SampleB.F.fq.gz\n", "Renaming ./3L_0_R1_.fastq.gz -> Pop3_SampleL.F.fq.gz\n", "Renaming ./1C_0_R1_.fastq.gz -> Pop1_SampleC.F.fq.gz\n", "Renaming ./1A_0_R1_.fastq.gz -> Pop1_SampleA.F.fq.gz\n", "Renaming ./2G_0_R1_.fastq.gz -> Pop2_SampleG.F.fq.gz\n", "Renaming ./2E_0_R1_.fastq.gz -> Pop2_SampleE.F.fq.gz\n", "Renaming ./3J_0_R1_.fastq.gz -> Pop3_SampleJ.F.fq.gz\n", "Renaming ./2F_0_R1_.fastq.gz -> Pop2_SampleF.F.fq.gz\n", "Renaming ./3I_0_R1_.fastq.gz -> Pop3_SampleI.F.fq.gz\n", "Renaming ./1D_0_R1_.fastq.gz -> Pop1_SampleD.F.fq.gz\n", "export LD_LIBRARY_PATH=/home/iovercast/manuscript-analysis/dDocent//freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; export PATH=/home/iovercast/manuscript-analysis/dDocent/:$PATH; dDocent simlarge-config.txt\n", "dDocent 2.24 \n", "\n", "Contact jpuritz@gmail.com with any problems \n", "\n", " \n", "Checking for required software\n", "\n", "All required software is installed!\n", "dDocent will require input during the assembly stage. Please wait until prompt says it is safe to move program to the background.\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "Removing the _1 character and replacing with /1 in the name of every sequence\n", "Trimming reads and simultaneously assembling reference sequences\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n" ] } ], "source": [ "import ipyrad as ip\n", "import subprocess\n", "import glob\n", "\n", "## Set up directory structures. change the force flag if you want to\n", "## blow everything away and restart\n", "# force = True\n", "force = \"\"\n", "DDOCENT_SIMOUT=os.path.join(DDOCENT_DIR, \"SIMDATA/\")\n", "if force and os.path.exists(DDOCENT_SIMOUT):\n", " shutil.rmtree(DDOCENT_SIMOUT)\n", "if not os.path.exists(DDOCENT_SIMOUT):\n", " os.makedirs(DDOCENT_SIMOUT)\n", "for outdir in [\"no/\", \"lo/\", \"hi/\", \"large/\"]:\n", " tmpdir = os.path.join(DDOCENT_SIMOUT, \"sim\"+outdir)\n", " if not os.path.exists(tmpdir):\n", " os.makedirs(tmpdir)\n", "\n", " \n", "## go to the pyrad simulated data output directory\n", "for dir in [\"no\", \"lo\", \"hi\", \"large\"]:\n", " print(\"Doing - {}\".format(dir))\n", " ## Set barcode and raw data files\n", " simdata_dir = os.path.join(SIMULATION_DATA_DIR, \"sim\"+dir)\n", " bcodes = os.path.join(simdata_dir, \"sim{}_barcodes.txt\".format(dir))\n", " fastqs = os.path.join(simdata_dir, \"sim{}_R1_.fastq.gz\".format(dir))\n", "\n", " simout_dir = os.path.join(DDOCENT_SIMOUT, \"sim\"+dir)\n", " if not os.path.exists(simout_dir):\n", " os.mkdir(simout_dir)\n", " os.chdir(simout_dir)\n", "\n", " ## dDocent doesn't do demultiplexing, so we'll use ipyrad for it real quick\n", " data = ip.Assembly(\"sim\"+dir)\n", " \n", " data.set_params('raw_fastq_path', fastqs)\n", " data.set_params('barcodes_path', bcodes)\n", " data.write_params(force=True)\n", " \n", " cmd = \"ipyrad -p params-sim{}.txt -s 1\".format(dir)\n", " print(cmd)\n", " !time $cmd\n", " \n", " ## Now we have to rename all the files in the way dDocent expects them:\n", " ## 1A_0_R1_.fastq.gz -> Pop1_Sample1.F.fq.gz\n", " ##\n", " ## Have to cd to the directory cuz we have to be in the data dir when we run dDocent anyway\n", " os.chdir(\"sim{}_fastqs\".format(dir))\n", " fq_files = glob.glob(\"./*.fastq.gz\")\n", " for f in fq_files:\n", " newname = \"Pop{}_Sample{}.F.fq.gz\".format(f[2], f[3])\n", " print(\"Renaming {} -> {}\".format(f, newname))\n", " os.rename(f, newname)\n", "\n", " ## Write out the config file for this run.\n", " ## Compacted the config file into one long line here to make it not take up so much room\n", " config_file = \"sim{}-config.txt\".format(dir)\n", " with open(config_file, 'w') as outfile:\n", " outfile.write('Number of Processors\\n40\\nMaximum Memory\\n0\\nTrimming\\nyes\\nAssembly?\\nyes\\nType_of_Assembly\\nSE\\nClustering_Similarity%\\n0.85\\nMapping_Reads?\\nyes\\nMapping_Match_Value\\n1\\nMapping_MisMatch_Value\\n3\\nMapping_GapOpen_Penalty\\n5\\nCalling_SNPs?\\nyes\\nEmail\\nwatdo@mailinator.com\\n')\n", "\n", " cmd = \"export LD_LIBRARY_PATH={}/freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; \".format(DDOCENT_DIR)\n", " cmd += \"export PATH={}:$PATH; time dDocent {}\".format(DDOCENT_DIR, config_file)\n", " print(cmd)\n", " #!$cmd\n", " \n", " ## You have to post-process the vcf files to decompose complex genotypes \n", " ## and remove indels\n", " fullvcf = simdata_dir + \"TotalRawSNPs.vcf\"\n", " filtvcf = simdata_dir + \"Final.recode.vcf\"\n", " for f in [fullvcf, filtvcf]:\n", " print(\"Finalizing - {}\".format(f))\n", " ## Naming the new outfiles as .snps.vcf\n", " outfile = simdata_dir + f.split(\"/\")[-1].split(\".vcf\")[0] + \".snps.vcf\"\n", " cmd = \"export PATH={}:$PATH; vcfallelicprimitives {} > ddoc-tmp.vcf\".format(DDOCENT_DIR, f)\n", " print(cmd)\n", " #!$cmd\n", " cmd = \"export PATH={}:$PATH; vcftools --vcf ddoc-tmp.vcf --remove-indels --recode --recode-INFO-all --out {}\".format(DDOCENT_DIR, outfile)\n", " #!$cmd\n", " !rm ddoc-tmp.vcf\n" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "# Empirical Analysis\n", "An empirical analysis was conducted on the published data set from Eaton & Ree (2013), \n", "available de-multiplexed in fastQ format at the ncbi sequence read archive (SRA072507). \n", "The data are already demultiplexed ipyrad step 1 and stacks `process_radtags`\n", "are skipped. Because Stacks and ipyrad/PyRAD use different quality filtering methods we \n", "employed filtering by ipyrad only. This way quality filtering does not affect the results \n", "of comparison between the programs. After trimming the barcode and adapter \n", "read lengths are 69bp. At .85 similarity this is equivalent to 10 base differences. \n" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## ipyrad\n", "Some representative runs to give you an expectation of how long things should take:\n", "\n", "Step 1 - Linking samples:\n", "```\n", "real 0m33.881s\n", "user 0m24.455s\n", "sys 0m1.877s\n", "```\n", "Step 2 - Filtering Raw Reads:\n", "```\n", "real 7m48.600s\n", "user 0m15.081s\n", "sys 0m5.252s\n", "```\n", "Step 3 - Clustering within samples (`-c 13`):\n", "```\n", "real 29m56.718s\n", "user 0m24.510s\n", "sys 0m1.098s\n", "```\n", "Step 3 - Clustering within samples (`-c 40`):\n", "```\n", "## There is some overhead associated with parallelization so speedup\n", "## isn't quite 2x as fast, but is still considerable. With larger\n", "## number of samples the speed gain of massive parallelization\n", "## is pronounced.\n", "\n", "real 18m59.515s\n", "user 0m23.856s\n", "sys 0m1.058s\n", "```\n", "\n", "All steps 1-7 with `-c 40`:\n", "```\n", "real 50m25.647s\n", "user 9m34.733s\n", "sys 0m20.489s\n", "```\n", "\n", "All steps 1-7 with `-c 13`:\n", "```\n", "real 69m26.015s\n", "user 9m1.946s\n", "sys 0m20.285s\n", "```\n", "\n" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "### ipyrad params file (params-example.txt)\n", "Next, we write out the ipyrad params file to the ipyrad working direcotory. " ] }, { "cell_type": "code", "execution_count": 198, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "------- ipyrad params file (v.0.3.15)-------------------------------------------\n", "REALDATA ## [0] [assembly_name]: Assembly name. Used to name output directories for assembly steps\n", "./REALDATA ## [1] [project_dir]: Project dir (made in curdir if not present)\n", " ## [2] [raw_fastq_path]: Location of raw non-demultiplexed fastq files\n", " ## [3] [barcodes_path]: Location of barcodes file\n", "../example_empirical_rad/*.gz ## [4] [sorted_fastq_path]: Location of demultiplexed/sorted fastq files\n", "denovo ## [5] [assembly_method]: Assembly method (denovo, reference, denovo+reference, denovo-reference)\n", " ## [6] [reference_sequence]: Location of reference sequence file\n", "rad ## [7] [datatype]: Datatype (see docs): rad, gbs, ddrad, etc.\n", "TGCAG, ## [8] [restriction_overhang]: Restriction overhang (cut1,) or (cut1, cut2)\n", "4 ## [9] [max_low_qual_bases]: Max low quality base calls (Q<20) in a read\n", "33 ## [10] [phred_Qscore_offset]: phred Q score offset (only alternative=64)\n", "6 ## [11] [mindepth_statistical]: Min depth for statistical base calling\n", "6 ## [12] [mindepth_majrule]: Min depth for majority-rule base calling\n", "10000 ## [13] [maxdepth]: Max cluster depth within samples\n", "0.85 ## [14] [clust_threshold]: Clustering threshold for de novo assembly\n", "0 ## [15] [max_barcode_mismatch]: Max number of allowable mismatches in barcodes\n", "1 ## [16] [filter_adapters]: Filter for adapters/primers (1 or 2=stricter)\n", "69 ## [17] [filter_min_trim_len]: Min length of reads after adapter trim\n", "2 ## [18] [max_alleles_consens]: Max alleles per site in consensus sequences\n", "3, 3 ## [19] [max_Ns_consens]: Max N's (uncalled bases) in consensus (R1, R2)\n", "3, 3 ## [20] [max_Hs_consens]: Max Hs (heterozygotes) in consensus (R1, R2)\n", "2 ## [21] [min_samples_locus]: Min # samples per locus for output\n", "100, 100 ## [22] [max_SNPs_locus]: Max # SNPs per locus (R1, R2)\n", "3, 3 ## [23] [max_Indels_locus]: Max # of indels per locus (R1, R2)\n", "3 ## [24] [max_shared_Hs_locus]: Max # heterozygous sites per locus (R1, R2)\n", "0, 0 ## [25] [edit_cutsites]: Edit cut-sites (R1, R2) (see docs)\n", "2, 2, 2, 2 ## [26] [trim_overhang]: Trim overhang (see docs) (R1>, , /@777V8?7X_NY[1,]_1$7T?_Z#OW!A1S/,U*WDA?KRF63,G-X_I7R,RLLK/\\nM2-6VJBUE4HO4*BT%3J@TM1!DK*A,%(_V<6\\\\.,,$!*JU^ HX!%?3?PLQQ4*_@\\nMF2!94EIK-Y<+*I6E2@LC2MN/=L(\\'@@0$FM^Q!3?VAE7_#F [!IM1VHKL>;<>N3,*NQ]YE8E2W:J=/\"?=ZBB$7:FL6Q_!\\nM0KV -\" M%D*+,A4#\"J8_?[,,-Y:#DG?JDM?^@AEQ4[O!=@+;#=1N\"%%V*MJ#\\nM_Q+X&;?PA!5;Y-^=+P_3J] 5?![EJL[\\nM=E.CF.;<\" .B+_R>8\"1GE\\'9-SDZHMMQ0[_)-,NX[<7&$D&?1=+J79^+Z1;72\\nM*+Q+DRHMF%HLC+!@\\\\E:Z)&^F8*:>*E$G/+=\"E\\\\C7K7A[>MR/3O<[-\\'%]H9!E\\nM)BJ[8@9ID<8ZF3B?<&1O]ZVGL_;@G2R7![$D79:\"_]1U+GYC@!V=SJZ\\';O4Q\\nMQV2,;P_\\'-.3&XS492?,:C5(W\\'\\'?2KD!H?$,PT3@^.]EW;M< / BS\"+M9J=P)\\nM\\\\UV Q<%H8_8>H(=ZK;=:CG:<><-QTM14T[I8(4W!;=KZ4-;%\\'\"Y ?XB%NN-S\\nMQ*;=@CX\\'A]J>%TT.8\\'-*AVIQ?,8S7N&_*]^_O25<,8UU5&E9X#_U)DXIR=N@\\nM)*%15Z_V\\\\[Q\\\\X$\\'B&7Y;L%R43=(Q6B(COJGPS!!?./J&FMSF*#G F;,F=(B,\\nM0$I9JDK<5ZT>&RM5@#32AOLM[*C-IH>9:#J@78KT3*WP+[9)+OY A.K2%2ID\\nMZ=M!_S%1[PH=Y@K]Y0\">9-SPG&_SG -H)1 =]6N]5/9YE@/BEC2B9XT4T,[2\\nMVC2I.6H%XN/F5WQ=A!>)4^* )CO$Z F2QHU_+KYUT.\\'&$3E3!_I_1:>1./N$\\nM3I)O Z.8I+<_B;X/^KB!-BONNN[Y-OI#[%WMH)J>I$%L=DG>,SF]BTQ:AEO\\'\\nM X\\'C ^;N%AH&Y!:OAG;B]TCI7X#7FG;MO ^A(_AK0FT#E1W#@\\nMQ8[3MRQ.VJ$:& H#C)2&M\\\\=_GRU$J0 \\n \\nend\\n'\n", "print(params.decode(\"uu\").decode(\"zlib\"))\n", "with open(os.path.join(IPYRAD_DIR, \"params-example.txt\"), 'w') as outfile:\n", " outfile.write(params.decode(\"uu\").decode(\"zlib\"))\n", "\n", "## If you want to make changes to the params file you can generate a new uuencoded string\n", "## by editing the pyrad/params-pyrad.txt file and running these lines. If you run this\n", "## at a terminal inside python shell then it'll print out nicely as one big fat line. \n", "#with open(os.path.join(PYRAD_DIR, \"params-pyrad.txt\")) as infile:\n", "# infile.read().encode(\"zlib\").encode(\"uu\")" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "### ipyrad - Import samples and filter raw data\n", "This step should take 5-10 minutes depending on the available resources." ] }, { "cell_type": "code", "execution_count": 76, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.3.36]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: REALDATA\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Linking sorted fastq data to Samples\n", " Linking to demultiplexed fastq files in: /home/iovercast/manuscript-analysis/example_empirical_rad/*.gz\n", " 13 new Samples created in 'REALDATA'.\n", " 13 fastq files linked to 13 new Samples.\n", "\n", " Step 2: Filtering reads \n", "\r", " [ ] 0% processing reads | 0:00:00 \r", " [ ] 0% processing reads | 0:00:01 \r", " [ ] 0% processing reads | 0:00:02 \r", " [ ] 0% processing reads | 0:00:03 \r", " [ ] 0% processing reads | 0:00:04 \r", " [ ] 0% processing reads | 0:00:05 \r", " [ ] 0% processing reads | 0:00:06 \r", " [ ] 0% processing reads | 0:00:07 \r", " [ ] 0% processing reads | 0:00:08 \r", " [ ] 0% processing reads | 0:00:09 \r", " [ ] 0% processing reads | 0:00:10 \r", " [ ] 0% processing reads | 0:00:11 \r", " [ ] 0% processing reads | 0:00:12 \r", " [ ] 0% processing reads | 0:00:13 \r", " [ ] 0% processing reads | 0:00:14 \r", " [ ] 0% processing reads | 0:00:15 \r", " [ ] 0% processing reads | 0:00:16 \r", " [ ] 0% processing reads | 0:00:17 \r", " [ ] 0% processing reads | 0:00:18 \r", " [ ] 0% processing 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params-example.txt -s 12" ] }, { "cell_type": "code", "execution_count": 199, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.3.36]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " loading Assembly: REALDATA\n", " from saved path: ~/manuscript-analysis/ipyrad/REALDATA/REALDATA.json\n", " local compute node: [40 cores] on node001\n", "\n", " Step 3: Clustering/Mapping reads\n", "\r", " [ ] 0% dereplicating | 0:00:00 \r", " [ ] 0% dereplicating | 0:00:01 \r", " [### ] 15% dereplicating | 0:00:02 \r", " [####### ] 38% dereplicating | 0:00:03 \r", " [############ ] 61% dereplicating | 0:00:04 \r", " [################ ] 84% dereplicating | 0:00:05 \r", " [####################] 100% dereplicating | 0:00:06 \r", " [####################] 100% dereplicating | 0:00:07 \n", 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" [################## ] 92% aligning | 0:08:54 \r", " [################## ] 92% aligning | 0:08:55 \r", " [################## ] 92% aligning | 0:08:56 \r", " [################## ] 92% aligning | 0:08:57 \r", " [################## ] 92% aligning | 0:08:58 \r", " [################## ] 92% aligning | 0:08:59 \r", " [################## ] 92% aligning | 0:09:00 \r", " [################## ] 92% aligning | 0:09:01 \r", " [####################] 100% aligning | 0:09:02 \r", " [####################] 100% aligning | 0:09:03 \n", "\r", " [################## ] 92% concatenating | 0:00:00 \r", " [################## ] 92% concatenating | 0:00:01 \r", " [################## ] 92% concatenating | 0:00:02 \r", " [################## ] 92% concatenating | 0:00:03 \r", " [################## ] 92% concatenating | 0:00:04 \r", " [################## ] 92% concatenating | 0:00:05 \r", " [################## ] 92% concatenating | 0:00:06 \r", " [################## ] 92% concatenating | 0:00:07 \r", " 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[H, E] | 0:00:13 \r", " [ ] 0% inferring [H, E] | 0:00:14 \r", " [ ] 0% inferring [H, E] | 0:00:14 \r", " [ ] 0% inferring [H, E] | 0:00:15 \r", " [ ] 0% inferring [H, E] | 0:00:15 \r", " [ ] 0% inferring [H, E] | 0:00:16 \r", " [ ] 0% inferring [H, E] | 0:00:16 \r", " [ ] 0% inferring [H, E] | 0:00:17 \r", " [ ] 0% inferring [H, E] | 0:00:17 \r", " [ ] 0% inferring [H, E] | 0:00:18 \r", " [ ] 0% inferring [H, E] | 0:00:18 \r", " [ ] 0% inferring [H, E] | 0:00:19 \r", " [ ] 0% inferring [H, E] | 0:00:19 \r", " [ ] 0% inferring [H, E] | 0:00:20 \r", " [ ] 0% inferring [H, E] | 0:00:20 \r", " [ ] 0% inferring [H, E] | 0:00:21 \r", " [ ] 0% inferring [H, E] | 0:00:21 \r", " [ ] 0% inferring [H, E] | 0:00:22 \r", " [ ] 0% inferring [H, E] | 0:00:22 \r", " [ ] 0% inferring [H, E] | 0:00:23 \r", " [ ] 0% inferring [H, E] | 0:00:23 \r", " [ ] 0% inferring [H, E] | 0:00:24 \r", " [ ] 0% inferring [H, E] | 0:00:24 \r", " [ ] 0% inferring [H, E] | 0:00:25 \r", " [ ] 0% inferring [H, E] | 0:00:25 \r", " [ ] 0% inferring [H, E] | 0:00:26 \r", " [ ] 0% inferring [H, E] | 0:00:26 \r", " [ ] 0% inferring [H, E] | 0:00:27 \r", " [ ] 0% inferring [H, E] | 0:00:27 \r", " [ ] 0% inferring [H, E] | 0:00:28 \r", " [ ] 0% inferring [H, E] | 0:00:28 \r", " [ ] 0% inferring [H, E] | 0:00:29 \r", " [ ] 0% inferring [H, E] | 0:00:29 \r", " [ ] 0% inferring [H, E] | 0:00:30 \r", " [ ] 0% inferring [H, E] | 0:00:30 \r", " [ ] 0% inferring [H, E] | 0:00:31 \r", " [ ] 0% inferring [H, E] | 0:00:31 \r", " [ ] 0% inferring [H, E] | 0:00:32 \r", " [ ] 0% inferring [H, E] | 0:00:32 \r", " [ ] 0% inferring [H, E] | 0:00:33 \r", " [ ] 0% inferring [H, E] | 0:00:33 \r", " [ ] 0% inferring [H, E] | 0:00:34 \r", " [ ] 0% inferring [H, E] | 0:00:34 \r", " [ ] 0% inferring [H, E] | 0:00:35 \r", " [ ] 0% inferring [H, E] | 0:00:35 \r", " [ ] 0% inferring [H, E] | 0:00:36 \r", " [ ] 0% inferring [H, E] | 0:00:36 \r", " [ ] 0% inferring [H, E] | 0:00:37 \r", " [ ] 0% inferring [H, E] | 0:00:37 \r", " [ ] 0% inferring [H, E] | 0:00:38 \r", " [ ] 0% inferring [H, E] | 0:00:38 \r", " [ ] 0% inferring [H, E] | 0:00:39 \r", " [ ] 0% inferring [H, E] | 0:00:39 \r", " [ ] 0% inferring [H, E] | 0:00:40 \r", " [ ] 0% inferring [H, E] | 0:00:40 \r", " [ ] 0% inferring [H, E] | 0:00:41 \r", " [ ] 0% inferring [H, E] | 0:00:41 \r", " [ ] 0% inferring [H, E] | 0:00:42 \r", " [ ] 0% inferring [H, E] | 0:00:42 \r", " [ ] 0% inferring [H, E] | 0:00:43 \r", " [ ] 0% inferring [H, E] | 0:00:43 \r", " [ ] 0% inferring [H, E] | 0:00:44 \r", " [ ] 0% inferring [H, E] | 0:00:44 \r", " [ ] 0% inferring [H, E] | 0:00:45 \r", " [ ] 0% inferring [H, E] | 0:00:45 \r", " [ ] 0% inferring [H, E] | 0:00:46 \r", " [ ] 0% inferring [H, E] | 0:00:46 \r", " [ ] 0% inferring [H, E] | 0:00:47 \r", " [ ] 0% inferring [H, E] | 0:00:47 \r", " [ ] 0% inferring [H, E] | 0:00:48 \r", " [ ] 0% inferring [H, E] | 0:00:48 \r", " [ ] 0% inferring [H, E] | 0:00:49 \r", " [ ] 0% inferring [H, E] | 0:00:49 \r", " [ ] 0% inferring [H, E] | 0:00:50 \r", " [ ] 0% inferring [H, E] | 0:00:50 \r", " [ ] 0% inferring [H, E] | 0:00:51 \r", " [ ] 0% inferring [H, E] | 0:00:51 \r", " [ ] 0% inferring [H, E] | 0:00:52 \r", " [ ] 0% inferring [H, E] | 0:00:52 \r", " [ ] 0% inferring [H, E] | 0:00:53 \r", " [ ] 0% inferring [H, E] | 0:00:53 \r", " [ ] 0% inferring [H, E] | 0:00:54 \r", " [ ] 0% inferring [H, E] | 0:00:54 \r", " [ ] 0% inferring [H, E] | 0:00:55 \r", " [ ] 0% inferring [H, E] | 0:00:55 \r", " [ ] 0% inferring [H, E] | 0:00:56 \r", " [ ] 0% inferring [H, E] | 0:00:56 \r", " [ ] 0% inferring [H, E] | 0:00:57 \r", " [ ] 0% inferring [H, E] | 0:00:57 \r", " [ ] 0% inferring [H, E] | 0:00:58 \r", " [ ] 0% inferring [H, E] | 0:00:58 \r", " [ ] 0% inferring [H, E] | 0:00:59 \r", " [ ] 0% inferring [H, E] | 0:00:59 \r", " [ ] 0% inferring [H, E] | 0:01:00 \r", " [# ] 7% inferring [H, E] | 0:01:00 \r", " [# ] 7% inferring [H, E] | 0:01:01 \r", " [# ] 7% inferring [H, E] | 0:01:01 \r", " [### ] 15% inferring [H, E] | 0:01:02 \r", " [### ] 15% inferring [H, E] | 0:01:02 \r", " [### ] 15% inferring [H, E] | 0:01:03 \r", " [### ] 15% inferring [H, E] | 0:01:03 \r", " [### ] 15% inferring [H, E] | 0:01:04 \r", " [### ] 15% inferring [H, E] | 0:01:04 \r", " [### ] 15% inferring [H, E] | 0:01:05 \r", " [### ] 15% inferring [H, E] | 0:01:05 \r", " [### ] 15% inferring [H, E] | 0:01:06 \r", " [### ] 15% inferring [H, E] | 0:01:06 \r", " [### ] 15% inferring [H, E] | 0:01:07 \r", " [### ] 15% inferring [H, E] | 0:01:07 \r", " [###### ] 30% inferring [H, E] | 0:01:08 \r", " [####### ] 38% inferring [H, E] | 0:01:08 \r", " [####### ] 38% inferring [H, E] | 0:01:09 \r", " [####### ] 38% inferring [H, E] | 0:01:09 \r", " [####### ] 38% inferring [H, E] | 0:01:10 \r", " [####### ] 38% inferring [H, E] | 0:01:10 \r", " [####### ] 38% inferring [H, E] | 0:01:11 \r", " [####### ] 38% inferring [H, E] | 0:01:11 \r", " [####### ] 38% inferring [H, E] | 0:01:12 \r", " [####### ] 38% inferring [H, E] | 0:01:12 \r", " [####### ] 38% inferring [H, E] | 0:01:13 \r", " [####### ] 38% inferring [H, E] | 0:01:13 \r", " [######### ] 46% inferring [H, E] | 0:01:14 \r", " [######### ] 46% inferring [H, E] | 0:01:14 \r", " [######### ] 46% inferring [H, E] | 0:01:15 \r", " [########## ] 53% inferring [H, E] | 0:01:15 \r", " [########## ] 53% inferring [H, E] | 0:01:16 \r", " [########## ] 53% inferring [H, E] | 0:01:16 \r", " [########## ] 53% inferring [H, E] | 0:01:17 \r", " [########## ] 53% inferring [H, E] | 0:01:17 \r", " [########## ] 53% inferring [H, E] | 0:01:18 \r", " [############ ] 61% inferring [H, E] | 0:01:18 \r", " [############ ] 61% inferring [H, E] | 0:01:19 \r", " [############ ] 61% inferring [H, E] | 0:01:19 \r", " [############ ] 61% inferring [H, E] | 0:01:20 \r", " [############ ] 61% inferring [H, E] | 0:01:20 \r", " [############ ] 61% inferring [H, E] | 0:01:21 \r", " [############ ] 61% inferring [H, E] | 0:01:21 \r", " [############ ] 61% inferring [H, E] | 0:01:22 \r", " [############ ] 61% inferring [H, E] | 0:01:22 \r", " [############ ] 61% inferring [H, E] | 0:01:23 \r", " [############ ] 61% inferring [H, E] | 0:01:23 \r", " [############ ] 61% inferring [H, E] | 0:01:24 \r", " [############ ] 61% inferring [H, E] | 0:01:24 \r", " [############ ] 61% inferring [H, E] | 0:01:25 \r", " [############# ] 69% inferring [H, E] | 0:01:25 \r", " [############# ] 69% inferring [H, E] | 0:01:26 \r", " [############# ] 69% inferring [H, E] | 0:01:26 \r", " [############# ] 69% inferring [H, E] | 0:01:27 \r", " [############# ] 69% inferring [H, E] | 0:01:27 \r", " [############# ] 69% inferring [H, E] | 0:01:28 \r", " [############# ] 69% inferring [H, E] | 0:01:28 \r", " [############# ] 69% inferring [H, E] | 0:01:29 \r", " [############### ] 76% inferring [H, E] | 0:01:29 \r", " [############### ] 76% inferring [H, E] | 0:01:30 \r", " [############### ] 76% inferring [H, E] | 0:01:30 \r", " [############### ] 76% inferring [H, E] | 0:01:31 \r", " [############### ] 76% inferring [H, E] | 0:01:31 \r", " [############### ] 76% inferring [H, E] | 0:01:32 \r", " [############### ] 76% inferring [H, E] | 0:01:32 \r", " [############### ] 76% inferring [H, E] | 0:01:33 \r", " [################ ] 84% inferring [H, E] | 0:01:33 \r", " [################ ] 84% inferring [H, E] | 0:01:34 \r", " [################## ] 92% inferring [H, E] | 0:01:34 \r", " [################## ] 92% inferring [H, E] | 0:01:35 \r", " [################## ] 92% inferring [H, E] | 0:01:35 \r", " [################## ] 92% inferring [H, E] | 0:01:36 \r", " [################## ] 92% inferring [H, E] | 0:01:36 \r", " [################## ] 92% inferring [H, E] | 0:01:37 \r", " [################## ] 92% inferring [H, E] | 0:01:37 \r", " [################## ] 92% inferring [H, E] | 0:01:38 \r", " [################## ] 92% inferring [H, E] | 0:01:38 \r", " [################## ] 92% inferring [H, E] | 0:01:39 \r", " [################## ] 92% inferring [H, E] | 0:01:39 \r", " [####################] 100% inferring [H, E] | 0:01:39 \n", "\n", " Step 5: Consensus base calling \n", " Mean error [0.00246 sd=0.00094]\n", " Mean hetero [0.01883 sd=0.00313]\n", "\r", " [ ] 0% consensus calling | 0:00:00 \r", " [ ] 0% consensus calling | 0:00:04 \r", " [ ] 0% consensus calling | 0:00:09 \r", " [ ] 0% consensus calling | 0:00:17 \r", " [ ] 0% consensus calling | 0:00:25 \r", " [ ] 0% consensus calling | 0:00:32 \r", " [ ] 0% consensus calling | 0:00:43 \r", " [ ] 0% consensus calling | 0:00:50 \r", " [ ] 0% consensus calling | 0:00:59 \r", " [ ] 0% consensus calling | 0:01:07 \r", " [ ] 0% consensus calling | 0:01:16 \r", " [ ] 0% consensus calling | 0:01:25 \r", " [ ] 0% consensus calling | 0:01:40 \r", " [ ] 0% consensus calling | 0:01:50 \r", " [################## ] 90% consensus calling | 0:01:51 \r", " [################## ] 90% consensus calling | 0:01:52 \r", " [################## ] 90% consensus calling | 0:01:53 \r", " [################## ] 90% consensus calling | 0:01:54 \r", " [################## ] 92% consensus calling | 0:01:55 \r", " [################## ] 94% consensus calling | 0:01:56 \r", " [################## ] 94% consensus calling | 0:01:57 \r", " [################### ] 95% consensus calling | 0:01:58 \r", " [################### ] 95% consensus calling | 0:01:59 \r", " [################### ] 95% consensus calling | 0:02:00 \r", " [################### ] 95% consensus calling | 0:02:01 \r", " [################### ] 95% consensus calling | 0:02:02 \r", " [################### ] 95% consensus calling | 0:02:03 \r", " [################### ] 95% consensus calling | 0:02:04 \r", " [################### ] 96% consensus calling | 0:02:05 \r", " [################### ] 96% consensus calling | 0:02:06 \r", " [################### ] 96% consensus calling | 0:02:07 \r", " [################### ] 97% consensus calling | 0:02:08 \r", " [################### ] 97% consensus calling | 0:02:09 \r", " [################### ] 97% consensus calling | 0:02:10 \r", " [################### ] 97% consensus calling | 0:02:11 \r", " [################### ] 99% consensus calling | 0:02:12 \r", " [################### ] 99% consensus calling | 0:02:13 \r", " [################### ] 99% consensus calling | 0:02:14 \r", " [################### ] 99% consensus calling | 0:02:15 \r", " [####################] 100% consensus calling | 0:02:16 \r", " [####################] 100% consensus calling | 0:02:16 \n", "\n", " Step 6: Clustering across 13 samples at 0.85 similarity\n", "\r", " [ ] 0% concat/shuffle input | 0:00:00 \r", " [ ] 0% concat/shuffle input | 0:00:00 \r", " [ ] 0% concat/shuffle input | 0:00:01 \r", " [ ] 0% concat/shuffle input | 0:00:01 \r", " [ ] 0% concat/shuffle input | 0:00:02 \r", " [ ] 0% concat/shuffle input | 0:00:02 \r", " [ ] 0% concat/shuffle input | 0:00:03 \r", " [ ] 0% concat/shuffle input | 0:00:03 \r", " [ ] 0% concat/shuffle input | 0:00:04 \r", " [ ] 0% concat/shuffle input | 0:00:04 \r", " [ ] 0% concat/shuffle input | 0:00:05 \r", " [ ] 0% concat/shuffle input | 0:00:05 \r", " [####################] 100% concat/shuffle input | 0:00:06 \n", "\r", " [ ] 0% clustering across | 0:00:00 \r", " [ ] 0% clustering across | 0:00:00 \r", " [ ] 0% clustering across | 0:00:00 \r", " [ ] 0% clustering across | 0:00:00 \r", " [ ] 0% clustering across | 0:00:00 \r", " [ ] 0% clustering across | 0:00:00 \r", " [ ] 0% clustering across | 0:00:00 \r", " [ ] 0% clustering across | 0:00:00 \r", " [ ] 0% clustering across | 0:00:00 \r", " [ ] 0% clustering across | 0:00:01 \r", " [ ] 0% clustering across | 0:00:01 \r", " [ ] 0% clustering across | 0:00:01 \r", " [ ] 0% clustering across | 0:00:01 \r", " [ ] 1% clustering across | 0:00:01 \r", " [ ] 1% clustering across | 0:00:01 \r", " [ ] 2% clustering across | 0:00:01 \r", " [ ] 2% clustering across | 0:00:01 \r", " [ ] 3% clustering across | 0:00:01 \r", " [ ] 3% clustering across | 0:00:01 \r", " [ ] 4% clustering across | 0:00:01 \r", " [ ] 4% clustering across | 0:00:01 \r", " [# ] 5% clustering across | 0:00:01 \r", " [# ] 5% clustering across | 0:00:01 \r", " [# ] 6% clustering across | 0:00:02 \r", " [# ] 6% clustering across | 0:00:02 \r", " [# ] 7% clustering across | 0:00:02 \r", " [# ] 7% clustering across | 0:00:02 \r", " [# ] 8% clustering across | 0:00:02 \r", " [# ] 8% clustering across | 0:00:03 \r", " [# ] 9% clustering across | 0:00:03 \r", " [# ] 9% clustering across | 0:00:03 \r", " [## ] 10% clustering across | 0:00:04 \r", " [## ] 10% clustering across | 0:00:04 \r", " [## ] 11% clustering across | 0:00:04 \r", " [## ] 11% clustering across | 0:00:05 \r", " [## ] 12% clustering across | 0:00:05 \r", " [## ] 12% clustering across | 0:00:05 \r", " [## ] 13% clustering across | 0:00:05 \r", " [## ] 13% clustering across | 0:00:05 \r", " [## ] 14% clustering across | 0:00:07 \r", " [## ] 14% clustering across | 0:00:07 \r", " [### ] 15% clustering across | 0:00:07 \r", " [### ] 15% clustering across | 0:00:08 \r", " [### ] 16% clustering across | 0:00:09 \r", " [### ] 16% clustering across | 0:00:10 \r", " [### ] 17% clustering across | 0:00:10 \r", " [### ] 17% clustering across | 0:00:11 \r", " [### ] 18% clustering across | 0:00:11 \r", " [### ] 18% clustering across | 0:00:12 \r", " [### ] 19% clustering across | 0:00:12 \r", " [### ] 19% clustering across | 0:00:12 \r", " [#### ] 20% clustering across | 0:00:13 \r", " [#### ] 20% clustering across | 0:00:13 \r", " [#### ] 21% clustering across | 0:00:13 \r", " [#### ] 21% clustering across | 0:00:13 \r", " [#### ] 22% clustering across | 0:00:14 \r", " [#### ] 22% clustering across | 0:00:14 \r", " [#### ] 23% clustering across | 0:00:14 \r", " [#### ] 23% clustering across | 0:00:14 \r", " [#### ] 24% clustering across | 0:00:15 \r", " [#### ] 24% clustering across | 0:00:16 \r", " [##### ] 25% clustering across | 0:00:16 \r", " [##### ] 25% clustering across | 0:00:17 \r", " [##### ] 26% clustering across | 0:00:17 \r", " [##### ] 26% clustering across | 0:00:17 \r", " [##### ] 27% clustering across | 0:00:19 \r", " [##### ] 27% clustering across | 0:00:19 \r", " [##### ] 28% clustering across | 0:00:19 \r", " [##### ] 28% clustering across | 0:00:19 \r", " [##### ] 28% clustering across | 0:00:19 \r", " [##### ] 28% clustering across | 0:00:20 \r", " [###### ] 30% clustering across | 0:00:20 \r", " [###### ] 30% clustering across | 0:00:21 \r", " [###### ] 31% clustering across | 0:00:22 \r", " [###### ] 31% clustering across | 0:00:22 \r", " [###### ] 32% clustering across | 0:00:23 \r", " [###### ] 32% clustering across | 0:00:23 \r", " [###### ] 33% clustering across | 0:00:24 \r", " [###### ] 33% clustering across | 0:00:24 \r", " [###### ] 34% clustering across | 0:00:24 \r", " [####### ] 35% clustering across | 0:00:26 \r", " [####### ] 35% clustering across | 0:00:26 \r", " [####### ] 36% clustering across | 0:00:26 \r", " [####### ] 36% clustering across | 0:00:27 \r", " [####### ] 37% clustering across | 0:00:28 \r", " [####### ] 37% clustering across | 0:00:28 \r", " [####### ] 38% clustering across | 0:00:29 \r", " [####### ] 38% clustering across | 0:00:29 \r", " [####### ] 39% clustering across | 0:00:30 \r", " [####### ] 39% clustering across | 0:00:30 \r", " [######## ] 40% clustering across | 0:00:31 \r", " [######## ] 40% clustering across | 0:00:31 \r", " [######## ] 41% clustering across | 0:00:32 \r", " [######## ] 41% clustering across | 0:00:32 \r", " [######## ] 42% clustering across | 0:00:33 \r", " [######## ] 42% clustering across | 0:00:34 \r", " [######## ] 43% clustering across | 0:00:34 \r", " [######## ] 43% clustering across | 0:00:35 \r", " [######## ] 44% clustering across | 0:00:36 \r", " [######## ] 44% clustering across | 0:00:37 \r", " [######### ] 45% clustering across | 0:00:37 \r", " [######### ] 45% clustering across | 0:00:37 \r", " [######### ] 46% clustering across | 0:00:38 \r", " [######### ] 46% clustering across | 0:00:39 \r", " [######### ] 47% clustering across | 0:00:40 \r", " [######### ] 47% clustering across | 0:00:41 \r", " [######### ] 48% clustering across | 0:00:42 \r", " [######### ] 48% clustering across | 0:00:42 \r", " [######### ] 49% clustering across | 0:00:42 \r", " [######### ] 49% clustering across | 0:00:43 \r", " [########## ] 50% clustering across | 0:00:43 \r", " [########## ] 50% clustering across | 0:00:43 \r", " [########## ] 51% clustering across | 0:00:43 \r", " [########## ] 51% clustering across | 0:00:43 \r", " [########## ] 52% clustering across | 0:00:44 \r", " [########## ] 52% clustering across | 0:00:45 \r", " [########## ] 53% clustering across | 0:00:46 \r", " [########## ] 53% clustering across | 0:00:46 \r", " [########## ] 54% clustering across | 0:00:47 \r", " [########## ] 54% clustering across | 0:00:47 \r", " [########### ] 55% clustering across | 0:00:48 \r", " [########### ] 55% clustering across | 0:00:49 \r", " [########### ] 56% clustering across | 0:00:49 \r", " [########### ] 56% clustering across | 0:00:50 \r", " [########### ] 56% clustering across | 0:00:52 \r", " [########### ] 56% clustering across | 0:00:52 \r", " [########### ] 57% clustering across | 0:00:53 \r", " [########### ] 57% clustering across | 0:00:54 \r", " [########### ] 59% clustering across | 0:00:54 \r", " [########### ] 59% clustering across | 0:00:56 \r", " [############ ] 60% clustering across | 0:00:57 \r", " [############ ] 60% clustering across | 0:00:57 \r", " [############ ] 61% clustering across | 0:00:58 \r", " [############ ] 61% clustering across | 0:00:59 \r", " [############ ] 62% clustering across | 0:01:01 \r", " [############ ] 62% clustering across | 0:01:02 \r", " [############ ] 63% clustering across | 0:01:02 \r", " [############ ] 63% clustering across | 0:01:03 \r", " [############ ] 64% clustering across | 0:01:04 \r", " [############ ] 64% clustering across | 0:01:04 \r", " [############# ] 65% clustering across | 0:01:04 \r", " [############# ] 65% clustering across | 0:01:06 \r", " [############# ] 66% clustering across | 0:01:06 \r", " [############# ] 66% clustering across | 0:01:07 \r", " [############# ] 67% clustering across | 0:01:07 \r", " [############# ] 67% clustering across | 0:01:08 \r", " [############# ] 68% clustering across | 0:01:09 \r", " [############# ] 69% clustering across | 0:01:09 \r", " [############# ] 69% clustering across | 0:01:10 \r", " [############## ] 70% clustering across | 0:01:10 \r", " [############## ] 70% clustering across | 0:01:11 \r", " [############## ] 71% clustering across | 0:01:12 \r", " [############## ] 71% clustering across | 0:01:13 \r", " [############## ] 72% clustering across | 0:01:14 \r", " [############## ] 72% clustering across | 0:01:14 \r", " [############## ] 73% clustering across | 0:01:16 \r", " [############## ] 73% clustering across | 0:01:16 \r", " [############## ] 74% clustering across | 0:01:17 \r", " [############## ] 74% clustering across | 0:01:17 \r", " [############### ] 75% clustering across | 0:01:17 \r", " [############### ] 75% clustering across | 0:01:18 \r", " [############### ] 76% clustering across | 0:01:19 \r", " [############### ] 76% clustering across | 0:01:21 \r", " [############### ] 77% clustering across | 0:01:22 \r", " [############### ] 77% clustering across | 0:01:22 \r", " [############### ] 78% clustering across | 0:01:22 \r", " [############### ] 78% clustering across | 0:01:22 \r", " [############### ] 79% clustering across | 0:01:23 \r", " [############### ] 79% clustering across | 0:01:24 \r", " [################ ] 80% clustering across | 0:01:24 \r", " [################ ] 80% clustering across | 0:01:24 \r", " [################ ] 81% clustering across | 0:01:25 \r", " [################ ] 81% clustering across | 0:01:25 \r", " [################ ] 82% clustering across | 0:01:26 \r", " [################ ] 82% clustering across | 0:01:26 \r", " [################ ] 83% clustering across | 0:01:27 \r", " 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0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 \r", " [# ] 5% building clusters | 0:00:05 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0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 \r", " [# ] 8% building clusters | 0:00:06 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clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 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clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 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clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 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8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 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0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 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8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 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clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 8% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 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clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 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9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 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9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 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clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 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clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 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clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 \r", " [# ] 9% building clusters | 0:00:07 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building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building clusters | 0:00:07 \r", " [## ] 10% building 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\r", " [############# ] 69% indexing clusters | 0:03:30 \r", " [############# ] 69% indexing clusters | 0:03:30 \r", " [############# ] 69% indexing clusters | 0:03:30 \r", " [############# ] 69% indexing clusters | 0:03:30 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:31 \r", " [############# ] 69% indexing clusters | 0:03:32 \r", " [############# ] 69% indexing clusters | 0:03:32 \r", " [############# ] 69% indexing clusters | 0:03:32 \r", " [############# ] 69% indexing clusters | 0:03:32 \r", " [############# ] 69% indexing clusters | 0:03:32 \r", " [############# ] 69% indexing clusters | 0:03:32 \r", " [############# ] 69% indexing clusters | 0:03:32 \r", " [############### ] 76% indexing clusters | 0:03:32 \r", " [############### ] 76% indexing clusters | 0:03:32 \r", " [############### ] 76% indexing clusters | 0:03:32 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:33 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:34 \r", " [############### ] 76% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:35 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:36 \r", " [################ ] 84% indexing clusters | 0:03:37 \r", " [################ ] 84% indexing clusters | 0:03:37 \r", " [################ ] 84% indexing clusters | 0:03:37 \r", " [################## ] 92% indexing clusters | 0:03:37 \r", " [################## ] 92% indexing clusters | 0:03:37 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[################## ] 92% building database | 0:09:06 \r", " [################## ] 92% building database | 0:09:06 \r", " [################## ] 92% building database | 0:09:06 \r", " [################## ] 92% building database | 0:09:06 \r", " [################## ] 92% building database | 0:09:06 \r", " [################## ] 92% building database | 0:09:06 \r", " [################## ] 92% building database | 0:09:06 \r", " [################## ] 92% building database | 0:09:06 \r", " [################## ] 92% building database | 0:09:06 \r", " [################## ] 92% building database | 0:09:07 \r", " [################## ] 92% building database | 0:09:07 \r", " [################## ] 92% building database | 0:09:07 \r", " [################## ] 92% building database | 0:09:07 \r", " [################## ] 92% building database | 0:09:07 \r", " [################## ] 92% building database | 0:09:07 \r", " [################## ] 92% building database | 0:09:07 \r", " [################## ] 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[################## ] 92% building database | 0:09:13 \r", " [################## ] 92% building database | 0:09:13 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:14 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 0:09:15 \r", " [################## ] 92% building database | 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[################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:16 \r", " [################## ] 92% building database | 0:09:17 \r", " [################## ] 92% building database | 0:09:17 \r", " [################## ] 92% building database | 0:09:17 \r", " [################## ] 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[################## ] 92% building database | 0:09:18 \r", " [################## ] 92% building database | 0:09:18 \r", " [################## ] 92% building database | 0:09:18 \r", " [################## ] 92% building database | 0:09:18 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:19 \r", " [################## ] 92% building database | 0:09:20 \r", " [################## ] 92% building database | 0:09:20 \r", " [################## ] 92% building database | 0:09:20 \r", " [################## ] 92% building database | 0:09:20 \r", " [################## ] 92% building database | 0:09:20 \r", " [################## ] 92% building database | 0:09:20 \r", " [################## ] 92% building database | 0:09:20 \r", " [################## ] 92% building database | 0:09:20 \r", " [################## ] 92% building database | 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[################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 92% building database | 0:09:26 \r", " [################## ] 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[################## ] 92% building database | 0:09:28 \r", " [################## ] 92% building database | 0:09:28 \r", " [################## ] 92% building database | 0:09:28 \r", " [################## ] 92% building database | 0:09:28 \r", " [################## ] 92% building database | 0:09:28 \r", " [################## ] 92% building database | 0:09:28 \r", " [################## ] 92% building database | 0:09:28 \r", " [################## ] 92% building database | 0:09:28 \r", " [################## ] 92% building database | 0:09:29 \r", " [################## ] 92% building database | 0:09:29 \r", " [################## ] 92% building database | 0:09:29 \r", " [################## ] 92% building database | 0:09:29 \r", " [################## ] 92% building database | 0:09:29 \r", " [################## ] 92% building database | 0:09:29 \r", " [################## ] 92% building database | 0:09:29 \r", " [################## ] 92% building database | 0:09:29 \r", " [################## ] 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[################## ] 92% building database | 0:09:33 \r", " [################## ] 92% building database | 0:09:33 \r", " [################## ] 92% building database | 0:09:33 \r", " [################## ] 92% building database | 0:09:33 \r", " [################## ] 92% building database | 0:09:33 \r", " [################## ] 92% building database | 0:09:33 \r", " [################## ] 92% building database | 0:09:33 \r", " [################## ] 92% building database | 0:09:33 \r", " [################## ] 92% building database | 0:09:34 \r", " [################## ] 92% building database | 0:09:34 \r", " [################## ] 92% building database | 0:09:34 \r", " [################## ] 92% building database | 0:09:34 \r", " [################## ] 92% building database | 0:09:34 \r", " [################## ] 92% building database | 0:09:34 \r", " [################## ] 92% building database | 0:09:34 \r", " [################## ] 92% building database | 0:09:34 \r", " [################## ] 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\r", " [####################] 100% building vcf file | 0:01:31 \r", " [####################] 100% building vcf file | 0:01:31 \n", "\r", " [#### ] 20% writing outfiles | 0:00:01 \r", " [######## ] 40% writing outfiles | 0:00:02 \r", " [############ ] 60% writing outfiles | 0:00:03 \r", " [############ ] 60% writing outfiles | 0:00:04 \r", " [############ ] 60% writing outfiles | 0:00:05 \r", " [################ ] 80% writing outfiles | 0:00:06 \r", " [################ ] 80% writing outfiles | 0:00:07 \r", " [################ ] 80% writing outfiles | 0:00:08 \r", " [################ ] 80% writing outfiles | 0:00:09 \r", " [################ ] 80% writing outfiles | 0:00:10 \r", " [################ ] 80% writing outfiles | 0:00:11 \r", " [################ ] 80% writing outfiles | 0:00:12 \r", " [####################] 100% writing outfiles | 0:00:13 \n", " Outfiles written to: ~/manuscript-analysis/ipyrad/REALDATA/REALDATA_outfiles\n", "\n" ] }, { "name": "stderr", "output_type": "stream", "text": [ "\n", "real\t44m47.378s\n", "user\t9m24.622s\n", "sys\t0m17.537s\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\" \"$IPYRAD_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"IPYRAD_DIR=$2\"\n", "cd $IPYRAD_DIR\n", "\n", "## Full run with 40 cores\n", "time ipyrad -p params-example.txt -s 34567 -f -c 40" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## pyRAD v.3\n", "In all cases with pyrad analysis we set `N processors` = 13.\n", "\n", "Step 2:\n", "```\n", "real\t1m37.142s\n", "user\t12m39.945s\n", "sys\t0m4.051s\n", "```\n", "\n", "Running steps 3-7:\n", "```\n", "real 250m58.693s\n", "user 1180m7.893s\n", "sys 12m1.399s\n", "```\n", "\n", "### pyRAD params file (params-pyrad.txt)\n" ] }, { "cell_type": "code", "execution_count": 99, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "==** parameter inputs for pyRAD version 3.0.63 **======================== affected step ==\n", "./REALDATA ## 1. Working directory (all)\n", " ## 2. Loc. of non-demultiplexed files (if not line 18) (s1)\n", " ## 3. Loc. of barcode file (if not line 18) (s1)\n", "vsearch ## 4. command (or path) to call vsearch (or usearch) (s3,s6)\n", "muscle ## 5. command (or path) to call muscle (s3,s7)\n", "TGCAG ## 6. Restriction overhang (e.g., C|TGCAG -> TGCAG) (s1,s2)\n", "13 ## 7. N processors (parallel) (all)\n", "6 ## 8. Mindepth: min coverage for a cluster (s4,s5)\n", "4 ## 9. NQual: max # sites with qual < 20 (or see line 20)(s2)\n", ".85 ## 10. Wclust: clustering threshold as a decimal (s3,s6)\n", "rad ## 11. Datatype: rad,gbs,pairgbs,pairddrad,(others:see docs)(all)\n", "2 ## 12. MinCov: min samples in a final locus (s7)\n", "3 ## 13. MaxSH: max inds with shared hetero site (s7)\n", "c85d6m2p3H3N3 ## 14. Prefix name for final output (no spaces) (s7)\n", "==== optional params below this line =================================== affected step ==\n", " ## 15.opt.: select subset (prefix* only selector) (s2-s7)\n", " ## 16.opt.: add-on (outgroup) taxa (list or prefix*) (s6,s7)\n", " ## 17.opt.: exclude taxa (list or prefix*) (s7)\n", "../ipyrad//REALDATA/REALDATA_edits/*_R1_* ## 18.opt.: loc. of de-multiplexed data (s2)\n", " ## 19.opt.: maxM: N mismatches in barcodes (def= 1) (s1)\n", " ## 20.opt.: phred Qscore offset (def= 33) (s2)\n", " ## 21.opt.: filter: def=0=NQual 1=NQual+adapters. 2=strict (s2)\n", " ## 22.opt.: a priori E,H (def= 0.001,0.01, if not estimated) (s5)\n", " ## 23.opt.: maxN: max Ns in a cons seq (def=5) (s5)\n", " ## 24.opt.: maxH: max heterozyg. sites in cons seq (def=5) (s5)\n", " ## 25.opt.: ploidy: max alleles in cons seq (def=2;see docs) (s4,s5)\n", " ## 26.opt.: maxSNPs: (def=100). Paired (def=100,100) (s7)\n", " ## 27.opt.: maxIndels: within-clust,across-clust (def. 3,99) (s3,s7)\n", " ## 28.opt.: random number seed (def. 112233) (s3,s6,s7)\n", " ## 29.opt.: trim overhang left,right on final loci, def(0,0) (s7)\n", " ## 30.opt.: output formats: p,n,a,s,v,u,t,m,k,g,* (see docs) (s7)\n", " ## 31.opt.: maj. base call at depth>x0&0Y8-PM.[CU=?KJ]^7,&!UZM7$(?PC])/\\nM0LZ!\"8T 2J\\\\._7S]&M*R# 8[6$D(7U0>@IJ!5/*$\\\\6I16E&7?(E5S43)#0R%\\nMNV>A%))#?!$@EHGW0*4;J$>J<\\\\5X [!G_799#NK9<%Q0[\\'_:LQ!R5554,A@Z\\nMCJDM K *4C@P[]^]\\nM+HR3^PLRSH@9!X.S/N1+K/C[@I8(2Y?P\"HRPJ*K?PA;P\"[^&=Y!$S0$8SKU(\\nMDB@8.B;\"B_%AV#A\"&S153;OBG\"-LH;DI5,F &BR;\\\\5Q4N,FZ8*\\\\+35D/,AKL\\nMFEIJ5S6? OZ4S!\\\\-J:G0W3MC[MNAL@5Z?>KJ9BHW@:/ZAGS[*A\\nM5>U,AI<4[2*QU%+E\"_.\"9A1):>\"1E^HWGI(P_JP/9>-V3.[F9 \\\\3XQ\"W#*>HIA+7@%D\\\\&HY%ULTC\\nMC$#)4_HOSZ#D3ES9/=B3%ILR=H*^\\'B(#.!/6G(X>[N*\\'T>$\"+MH\"\\nMRC;$&3_9;@8,\\';%_<>/5GB>[;(%1F5^GF%Z5,!6U>>%EW_8*3#+&9QG$P3;P\\nM3I_9!DZB%K@NG,:_FUQICI7/F@-OT-)T;RP>J3B)6V#L7^B;*3BL*&M2#&+_\\nM_H8R6N--$T*2^:C_\\'\\\\!))R \\\\4*&T@(_DIBTU\"J,H)O@W)M!V3&PA&%ZH^P\"!\\nMQ[W Z8;C6Y\\\\!MVVHY$H:%/POO\\\\WX)1_\\'@,\\\\VP&VX^\"SYLYJ\\';90WS6)GDV/ \\nMG4_K4@FV\\\\M!-(]N\\'F+Q=A^RZZ?1@3S9%W]]^,U,/$D=1@%F&^8UJZ;XA[MLM\\nM-GI-FIQO@#]CLRP1VL6LD\"=-\"R(TU\\\\H8_Z\\'9(H247%X&Z[&A![OSG\\\\:!1%4@\\nM%]4C;YHB:Y\\'B.$EV!.T[VC\\'LSH*HTVHSCI1\\\\9HD6\\\\P(C1FZZCR!.\\\\,.(1($G\\nMY/DPOO=,N&V*MH_H06^>-&8&F#AM7,W1J.47<5I_)-,KJ_[@;67N\"U\\nM#86\\\\YIK7FS[!W,>VV[_+,%YS50MN_&[Q4>#.AC@]0R%PXL,16B#3VS-5PV\\\\6\\nM$1\"KXWAH%35/&ZPX>\\\\T0_C7@_U\"%8@2B3*HUL)_1>K [\\nM&W;COZ/;#7[N &L\\\\M9_JL=UH\\\\A8VDMM@_S6:8,MS7;;IY*=Y29%4:HR8RXI+\\n/NSNQ;&EX=U89_ >8=H2-\\n \\nend\\n'\n", "print(params.decode(\"uu\").decode(\"zlib\"))\n", "with open(os.path.join(PYRAD_DIR, \"params-pyrad.txt\"), 'w') as outfile:\n", " outfile.write(params.decode(\"uu\").decode(\"zlib\"))\n", "\n", "## If you want to make changes to the params file you can generate a new uuencoded string\n", "## by editing the pyrad/params-pyrad.txt file and running these lines. If you run this\n", "## at a terminal inside python shell then it'll print out nicely as one big fat line. \n", "#with open(os.path.join(PYRAD_DIR, \"params-pyrad.txt\")) as infile:\n", "# infile.read().encode(\"zlib\").encode(\"uu\")" ] }, { "cell_type": "code", "execution_count": 98, "metadata": { "collapsed": true }, "outputs": [ { "name": "stderr", "output_type": "stream", "text": [ "\n", "\n", " ------------------------------------------------------------\n", " pyRAD : RADseq for phylogenetics & introgression analyses\n", " ------------------------------------------------------------\n", "\n", "\tsorted .fastq from /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/*_R1_* being used\n", "\tstep 2: editing raw reads \n", "\t.............\n", "real\t1m37.142s\n", "user\t12m39.945s\n", "sys\t0m4.051s\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\" \"$PYRAD_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"PYRAD_DIR=$2\"\n", "cd $PYRAD_DIR\n", "\n", "## Initial filtering\n", "##\n", "## pyrad v.3 performance is not improved by increasing the number\n", "## of cores available beyond the number of samples in the dataset\n", "## so results here are representative of increased values for\n", "## param 7 - N Processors (Parallel)\n", "time pyrad -p params-pyrad.txt -s 2" ] }, { "cell_type": "code", "execution_count": 197, "metadata": { "collapsed": false, "scrolled": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "\n", "\tfinished clustering\n", "\tingroup 29154_superba_R1_,30556_thamno_R1_,30686_cyathophylla_R1_,32082_przewalskii_R1_,33413_thamno_R1_,33588_przewalskii_R1_,35236_rex_R1_,35855_rex_R1_,38362_rex_R1_,39618_rex_R1_,40578_rex_R1_,41478_cyathophylloides_R1_,41954_cyathophylloides_R1_\n", "\taddon \n", "\texclude \n", "\t\n", "\tfinal stats written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/REALDATA/stats/c85d6m2p3H3N3.stats\n", "\toutput files being written to:\n", "\t /home/iovercast/manuscript-analysis/pyrad/REALDATA/outfiles/ directory\n", "\n" ] }, { "name": "stderr", "output_type": "stream", "text": [ "\n", "\n", " ------------------------------------------------------------\n", " pyRAD : RADseq for phylogenetics & introgression analyses\n", " ------------------------------------------------------------\n", "\n", "\n", "\tde-replicating files for clustering...\n", "\n", "\tstep 3: within-sample clustering of 13 samples at \n", "\t '.85' similarity. Running 13 parallel jobs\n", "\t \twith up to 6 threads per job. If needed, \n", "\t\tadjust to avoid CPU and MEM limits\n", "\n", "\tsample 29154_superba_R1_ finished, 117743 loci\n", "\tsample 33413_thamno_R1_ finished, 149210 loci\n", "\tsample 39618_rex_R1_ finished, 131379 loci\n", "\tsample 32082_przewalskii_R1_ finished, 132189 loci\n", "\tsample 33588_przewalskii_R1_ finished, 139491 loci\n", "\tsample 38362_rex_R1_ finished, 118064 loci\n", "\tsample 30556_thamno_R1_ finished, 186957 loci\n", "\tsample 30686_cyathophylla_R1_ finished, 203841 loci\n", "\tsample 41478_cyathophylloides_R1_ finished, 152744 loci\n", "\tsample 40578_rex_R1_ finished, 198186 loci\n", "\tsample 35855_rex_R1_ finished, 155901 loci\n", "\tsample 41954_cyathophylloides_R1_ finished, 262357 loci\n", "\tsample 35236_rex_R1_ finished, 373452 loci\n", "\n", "\tstep 4: estimating error rate and heterozygosity\n", "\t.............\n", "\tstep 5: creating consensus seqs for 13 samples, using H=0.01411 E=0.00200\n", "\t.............\n", "\tstep 6: clustering across 13 samples at '.85' similarity \n", "\n", "vsearch v2.0.3_linux_x86_64, 125.9GB RAM, 40 cores\n", "https://github.com/torognes/vsearch\n", "\n", "Reading file /home/iovercast/manuscript-analysis/pyrad/REALDATA/clust.85/cat.haplos_ 100%\n", "41107098 nt in 555794 seqs, min 62, max 77, avg 74\n", "Counting unique k-mers 100%\n", "Clustering 100%\n", "Sorting clusters 100%\n", "Writing clusters 100%\n", "Clusters: 180557 Size min 1, max 25, avg 3.1\n", "Singletons: 99736, 17.9% of seqs, 55.2% of clusters\n", ".............\n", "real\t274m33.324s\n", "user\t1523m51.077s\n", "sys\t123m20.369s\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\" \"$PYRAD_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"PYRAD_DIR=$2\"\n", "cd $PYRAD_DIR\n", "\n", "## All steps 3-7. We do not include step 2 because we are using\n", "## the ipyrad filtered data\n", "time pyrad -p params-pyrad.txt -s 34567" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## Stacks empirical analysis\n", "\n", "Runtime expectations:\n", "\n", "Ungapped with all flags set as below and 13 cores runtime exceeded 12 hours.\n", "\n", "Gapped alignment with parameters set as below and 40 cores:\n", "```\n", "real\t466m45.829s\n", "user\t17063m15.498s\n", "sys\t 4m24.564s\n", "```\n", "\n", "Ungapped all default assembly parameters:\n", "```\n", "real\t14m49.323s\n", "user\t20m29.222s\n", "sys\t 0m49.756s\n", "```\n", "\n", "Explanantions of all the stacks parameters. These settings are (close to being) analagous to the .85 clust similarity used in PyRAD given that read length are 100 bp.\n", "* -m 2: Minimum depth to make a stack\n", "* -M 10: Number of mismatches per locus within individuals\n", "* -N 10: Number of allowed mismatches in secondary reads\n", "* -n 10: Number of mismatches when clustering across individuals\n", "\n", "Housekeeping flags:\n", "* -T 40: Number of threads \n", "* -b 1: Batch id (Can be anything)\n", "* -S: Disable sql\n", "\n", "Populations flags:\n", "* -m 6: Minimum stack depth per locus when clustering across samples\n", "ustacks flags:\n", "* --max_locus_stacks 2: \n" ] }, { "cell_type": "code", "execution_count": 182, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "denovo_map.pl -m 2 -M 10 -N 10 -n 10 -T 13 -b 1 -S -d -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -X 'populations:-m 6' -X 'ustacks:--max_locus_stacks 2' -o /home/iovercast/manuscript-analysis/stacks/REALDATA/ungapped/ -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30556_thamno_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/32082_przewalskii_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/29154_superba_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/40578_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33588_przewalskii_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33413_thamno_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41478_cyathophylloides_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35855_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/39618_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/38362_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35236_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41954_cyathophylloides_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30686_cyathophylla_R1_.fastq \n" ] } ], "source": [ "## We build the stacks command w/ python because it'd be a pain\n", "## to do what the glob() call is doing in bash. Then we call\n", "## the command inside a bash cell because denovo_map expects\n", "## all the submodules to be in the PATH\n", "\n", "## Read in the filtered fastq from ipyrad step 2 \n", "IPYRAD_EDITS_DIR = os.path.join(IPYRAD_OUTPUT, \"REALDATA_edits/\")\n", "infiles = [\"-s \"+ff+\" \" for ff in glob.glob(IPYRAD_EDITS_DIR+\"*_R1_*\")]\n", "\n", "## Toggle the dryrun flag for testing\n", "DRYRUN=\"\"\n", "DRYRUN=\"-d\"\n", "\n", "OUTPUT_FORMATS = \"--vcf --genepop --structure --phase --fastphase --phylip \"\n", "cmd = \"denovo_map.pl -m 2 -M 10 -N 10 -n 10 -T 40 -b 1 -S \" + DRYRUN\\\n", " + \" -X \\'populations:\" + OUTPUT_FORMATS + \"\\'\"\\\n", " + \" -X \\'populations:-m 6\\' -X \\'ustacks:--max_locus_stacks 2\\' \"\\\n", " + \" -o \" + STACKS_UNGAP_OUT + \" \" + \"\".join(infiles)\n", "print(cmd)" ] }, { "cell_type": "code", "execution_count": 179, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "/home/iovercast/opt/bin/denovo_map.pl\n", "Identifying unique stacks; file 1 of 13 [30556_thamno_R1_]\n", "Identifying unique stacks; file 2 of 13 [32082_przewalskii_R1_]\n", "Identifying unique stacks; file 3 of 13 [29154_superba_R1_]\n", "Identifying unique stacks; file 4 of 13 [40578_rex_R1_]\n", "Identifying unique stacks; file 5 of 13 [33588_przewalskii_R1_]\n", "Identifying unique stacks; file 6 of 13 [33413_thamno_R1_]\n", "Identifying unique stacks; file 7 of 13 [41478_cyathophylloides_R1_]\n", "Identifying unique stacks; file 8 of 13 [35855_rex_R1_]\n", "Identifying unique stacks; file 9 of 13 [39618_rex_R1_]\n", "Identifying unique stacks; file 10 of 13 [38362_rex_R1_]\n", "Identifying unique stacks; file 11 of 13 [35236_rex_R1_]\n", "Identifying unique stacks; file 12 of 13 [41954_cyathophylloides_R1_]\n", "Identifying unique stacks; file 13 of 13 [30686_cyathophylla_R1_]\n" ] }, { "name": "stderr", "output_type": "stream", "text": [ "Found 13 sample file(s).\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30556_thamno_R1_.fastq -o REALDATA -i 1 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/32082_przewalskii_R1_.fastq -o REALDATA -i 2 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/29154_superba_R1_.fastq -o REALDATA -i 3 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/40578_rex_R1_.fastq -o REALDATA -i 4 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33588_przewalskii_R1_.fastq -o REALDATA -i 5 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33413_thamno_R1_.fastq -o REALDATA -i 6 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41478_cyathophylloides_R1_.fastq -o REALDATA -i 7 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35855_rex_R1_.fastq -o REALDATA -i 8 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/39618_rex_R1_.fastq -o REALDATA -i 9 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/38362_rex_R1_.fastq -o REALDATA -i 10 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35236_rex_R1_.fastq -o REALDATA -i 11 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41954_cyathophylloides_R1_.fastq -o REALDATA -i 12 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30686_cyathophylla_R1_.fastq -o REALDATA -i 13 -r -m 2 -M 10 -N 10 -p 13 --max_locus_stacks 2 2>&1\n", "\n", "Depths of Coverage for Processed Samples:\n", "\n", "Generating catalog...\n", " /home/iovercast/opt/bin/cstacks -b 1 -o REALDATA -s REALDATA/30556_thamno_R1_ -s REALDATA/32082_przewalskii_R1_ -s REALDATA/29154_superba_R1_ -s REALDATA/40578_rex_R1_ -s REALDATA/33588_przewalskii_R1_ -s REALDATA/33413_thamno_R1_ -s REALDATA/41478_cyathophylloides_R1_ -s REALDATA/35855_rex_R1_ -s REALDATA/39618_rex_R1_ -s REALDATA/38362_rex_R1_ -s REALDATA/35236_rex_R1_ -s REALDATA/41954_cyathophylloides_R1_ -s REALDATA/30686_cyathophylla_R1_ -n 10 -p 13 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/opt/bin/sstacks -b 1 -c REALDATA/batch_1 -o REALDATA -s REALDATA/30556_thamno_R1_ -s REALDATA/32082_przewalskii_R1_ -s REALDATA/29154_superba_R1_ -s REALDATA/40578_rex_R1_ -s REALDATA/33588_przewalskii_R1_ -s REALDATA/33413_thamno_R1_ -s REALDATA/41478_cyathophylloides_R1_ -s REALDATA/35855_rex_R1_ -s REALDATA/39618_rex_R1_ -s REALDATA/38362_rex_R1_ -s REALDATA/35236_rex_R1_ -s REALDATA/41954_cyathophylloides_R1_ -s REALDATA/30686_cyathophylla_R1_ -p 13 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/opt/bin/populations -b 1 -P REALDATA -s -t 13 --vcf --genepop --structure --phase --fastphase --phylip -m 6 2>&1\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\" \"$STACKS_DIR\" \"$cmd\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"STACKS_DIR=$2\"; export \"cmd=$3\"\n", "\n", "## We have to play a little cat and mouse game here because of quoting in some of the args\n", "## and how weird bash is we have to write the cmd to a file and then exec it.\n", "## If you try to just run $cmd it truncates the command at the first single tic. Hassle.\n", "cd $STACKS_DIR\n", "echo $cmd > stacks.sh; chmod 777 stacks.sh\n", "time ./stacks.sh" ] }, { "cell_type": "code", "execution_count": 200, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "denovo_map.pl -m 2 -M 10 -N 10 -n 10 -T 40 -b 1 -S --gapped -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -X 'populations:-m 6' -X 'ustacks:--max_locus_stacks 2' -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/ -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30556_thamno_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/32082_przewalskii_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/29154_superba_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/40578_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33588_przewalskii_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33413_thamno_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41478_cyathophylloides_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35855_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/39618_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/38362_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35236_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41954_cyathophylloides_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30686_cyathophylla_R1_.fastq \n" ] } ], "source": [ "## Do Gapped alignment\n", "\n", "## Read in the filtered fastq from ipyrad step 2 \n", "IPYRAD_EDITS_DIR = os.path.join(IPYRAD_OUTPUT, \"REALDATA_edits/\")\n", "infiles = [\"-s \"+ff+\" \" for ff in glob.glob(IPYRAD_EDITS_DIR+\"*_R1_*\")]\n", "\n", "## Toggle the dryrun flag for testing\n", "DRYRUN=\"\"\n", "#DRYRUN=\" -d \"\n", "\n", "OUTPUT_FORMATS = \"--vcf --genepop --structure --phase --fastphase --phylip \"\n", "cmd = \"denovo_map.pl -m 2 -M 10 -N 10 -n 10 -T 40 -b 1 -S --gapped \" + DRYRUN\\\n", " + \" -X \\'populations:\" + OUTPUT_FORMATS + \"\\'\"\\\n", " + \" -X \\'populations:-m 6\\' -X \\'ustacks:--max_locus_stacks 2\\' \"\\\n", " + \" -o \" + STACKS_GAP_OUT + \" \" + \"\".join(infiles)\n", "print(cmd)" ] }, { "cell_type": "code", "execution_count": 201, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "Identifying unique stacks; file 1 of 13 [30556_thamno_R1_]\n", "Identifying unique stacks; file 2 of 13 [32082_przewalskii_R1_]\n", "Identifying unique stacks; file 3 of 13 [29154_superba_R1_]\n", "Identifying unique stacks; file 4 of 13 [40578_rex_R1_]\n", "Identifying unique stacks; file 5 of 13 [33588_przewalskii_R1_]\n", "Identifying unique stacks; file 6 of 13 [33413_thamno_R1_]\n", "Identifying unique stacks; file 7 of 13 [41478_cyathophylloides_R1_]\n", "Identifying unique stacks; file 8 of 13 [35855_rex_R1_]\n", "Identifying unique stacks; file 9 of 13 [39618_rex_R1_]\n", "Identifying unique stacks; file 10 of 13 [38362_rex_R1_]\n", "Identifying unique stacks; file 11 of 13 [35236_rex_R1_]\n", "Identifying unique stacks; file 12 of 13 [41954_cyathophylloides_R1_]\n", "Identifying unique stacks; file 13 of 13 [30686_cyathophylla_R1_]\n" ] }, { "name": "stderr", "output_type": "stream", "text": [ "Found 13 sample file(s).\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30556_thamno_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 1 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/32082_przewalskii_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 2 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/29154_superba_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 3 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/40578_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 4 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33588_przewalskii_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 5 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33413_thamno_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 6 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41478_cyathophylloides_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 7 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35855_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 8 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/39618_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 9 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/38362_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 10 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35236_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 11 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41954_cyathophylloides_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 12 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30686_cyathophylla_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -i 13 -r -m 2 -M 10 -N 10 -p 40 --gapped --max_locus_stacks 2 2>&1\n", "\n", "Depths of Coverage for Processed Samples:\n", "30556_thamno_R1_: 9.02159x\n", "32082_przewalskii_R1_: 6.56423x\n", "29154_superba_R1_: 6.52959x\n", "40578_rex_R1_: 10.6561x\n", "33588_przewalskii_R1_: 7.22003x\n", "33413_thamno_R1_: 5.43012x\n", "41478_cyathophylloides_R1_: 17.5256x\n", "35855_rex_R1_: 9.8752x\n", "39618_rex_R1_: 7.46542x\n", "38362_rex_R1_: 12.7599x\n", "35236_rex_R1_: 7.90909x\n", "41954_cyathophylloides_R1_: 8.81844x\n", "30686_cyathophylla_R1_: 7.8596x\n", "\n", "Generating catalog...\n", " /home/iovercast/opt/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/30556_thamno_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/32082_przewalskii_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/29154_superba_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/40578_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/33588_przewalskii_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/33413_thamno_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/41478_cyathophylloides_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/35855_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/39618_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/38362_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/35236_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/41954_cyathophylloides_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/30686_cyathophylla_R1_ -n 10 -p 40 --gapped 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/opt/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/batch_1 -o /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/30556_thamno_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/32082_przewalskii_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/29154_superba_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/40578_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/33588_przewalskii_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/33413_thamno_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/41478_cyathophylloides_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/35855_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/39618_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/38362_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/35236_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/41954_cyathophylloides_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped/30686_cyathophylla_R1_ -p 40 --gapped 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/opt/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/REALDATA/gapped -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip -m 6 2>&1\n", "\n", "real\t456m59.593s\n", "user\t16632m17.688s\n", "sys\t3m1.172s\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\" \"$STACKS_DIR\" \"$cmd\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"STACKS_DIR=$2\"; export \"cmd=$3\"\n", "\n", "## Run the gapped alignment\n", "cd $STACKS_DIR\n", "echo $cmd > stacks.sh; chmod 777 stacks.sh\n", "time ./stacks.sh" ] }, { "cell_type": "code", "execution_count": 189, "metadata": { "collapsed": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "denovo_map.pl -T 40 -b 1 -S -X 'populations:--vcf --genepop --structure --phase --fastphase --phylip ' -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default/ -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30556_thamno_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/32082_przewalskii_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/29154_superba_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/40578_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33588_przewalskii_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33413_thamno_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41478_cyathophylloides_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35855_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/39618_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/38362_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35236_rex_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41954_cyathophylloides_R1_.fastq -s /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30686_cyathophylla_R1_.fastq \n" ] } ], "source": [ "## Do all default settings\n", "IPYRAD_EDITS_DIR = os.path.join(IPYRAD_OUTPUT, \"REALDATA_edits/\")\n", "infiles = [\"-s \"+ff+\" \" for ff in glob.glob(IPYRAD_EDITS_DIR+\"*_R1_*\")]\n", "\n", "## Toggle the dryrun flag for testing\n", "DRYRUN=\"\"\n", "#DRYRUN=\"-d\"\n", "\n", "OUTPUT_FORMATS = \"--vcf --genepop --structure --phase --fastphase --phylip \"\n", "cmd = \"denovo_map.pl -T 40 -b 1 -S \" + DRYRUN\\\n", " + \" -X \\'populations:\" + OUTPUT_FORMATS + \"\\'\"\\\n", " + \" -o \" + STACKS_DEFAULT_OUT + \" \" + \"\".join(infiles)\n", "print(cmd)" ] }, { "cell_type": "code", "execution_count": 190, "metadata": { "collapsed": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "Identifying unique stacks; file 1 of 13 [30556_thamno_R1_]\n", "Identifying unique stacks; file 2 of 13 [32082_przewalskii_R1_]\n", "Identifying unique stacks; file 3 of 13 [29154_superba_R1_]\n", "Identifying unique stacks; file 4 of 13 [40578_rex_R1_]\n", "Identifying unique stacks; file 5 of 13 [33588_przewalskii_R1_]\n", "Identifying unique stacks; file 6 of 13 [33413_thamno_R1_]\n", "Identifying unique stacks; file 7 of 13 [41478_cyathophylloides_R1_]\n", "Identifying unique stacks; file 8 of 13 [35855_rex_R1_]\n", "Identifying unique stacks; file 9 of 13 [39618_rex_R1_]\n", "Identifying unique stacks; file 10 of 13 [38362_rex_R1_]\n", "Identifying unique stacks; file 11 of 13 [35236_rex_R1_]\n", "Identifying unique stacks; file 12 of 13 [41954_cyathophylloides_R1_]\n", "Identifying unique stacks; file 13 of 13 [30686_cyathophylla_R1_]\n" ] }, { "name": "stderr", "output_type": "stream", "text": [ "Found 13 sample file(s).\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30556_thamno_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 1 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/32082_przewalskii_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 2 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/29154_superba_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 3 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/40578_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 4 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33588_przewalskii_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 5 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/33413_thamno_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 6 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41478_cyathophylloides_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 7 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35855_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 8 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/39618_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 9 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/38362_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 10 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/35236_rex_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 11 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/41954_cyathophylloides_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 12 -r -p 40 2>&1\n", " /home/iovercast/opt/bin/ustacks -t fastq -f /home/iovercast/manuscript-analysis/ipyrad/REALDATA/REALDATA_edits/30686_cyathophylla_R1_.fastq -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -i 13 -r -p 40 2>&1\n", "\n", "Depths of Coverage for Processed Samples:\n", "30556_thamno_R1_: 12.0867x\n", "32082_przewalskii_R1_: 8.05039x\n", "29154_superba_R1_: 7.86756x\n", "40578_rex_R1_: 13.8566x\n", "33588_przewalskii_R1_: 8.89649x\n", "33413_thamno_R1_: 6.77665x\n", "41478_cyathophylloides_R1_: 22.3755x\n", "35855_rex_R1_: 11.9406x\n", "39618_rex_R1_: 9.01107x\n", "38362_rex_R1_: 14.8266x\n", "35236_rex_R1_: 12.683x\n", "41954_cyathophylloides_R1_: 12.0704x\n", "30686_cyathophylla_R1_: 10.3671x\n", "\n", "Generating catalog...\n", " /home/iovercast/opt/bin/cstacks -b 1 -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/30556_thamno_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/32082_przewalskii_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/29154_superba_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/40578_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/33588_przewalskii_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/33413_thamno_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/41478_cyathophylloides_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/35855_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/39618_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/38362_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/35236_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/41954_cyathophylloides_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/30686_cyathophylla_R1_ -p 40 2>&1\n", "Matching samples to the catalog...\n", " /home/iovercast/opt/bin/sstacks -b 1 -c /home/iovercast/manuscript-analysis/stacks/REALDATA/default/batch_1 -o /home/iovercast/manuscript-analysis/stacks/REALDATA/default -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/30556_thamno_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/32082_przewalskii_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/29154_superba_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/40578_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/33588_przewalskii_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/33413_thamno_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/41478_cyathophylloides_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/35855_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/39618_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/38362_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/35236_rex_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/41954_cyathophylloides_R1_ -s /home/iovercast/manuscript-analysis/stacks/REALDATA/default/30686_cyathophylla_R1_ -p 40 2>&1\n", "Calculating population-level summary statistics\n", "/home/iovercast/opt/bin/populations -b 1 -P /home/iovercast/manuscript-analysis/stacks/REALDATA/default -s -t 40 --vcf --genepop --structure --phase --fastphase --phylip 2>&1\n", "\n", "real\t14m49.323s\n", "user\t20m29.222s\n", "sys\t0m49.756s\n" ] } ], "source": [ "%%bash -s \"$WORK_DIR\" \"$STACKS_DIR\" \"$cmd\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"STACKS_DIR=$2\"; export \"cmd=$3\"\n", "\n", "## Run with all defaults\n", "cd $STACKS_DIR\n", "echo $cmd > stacks.sh; chmod 777 stacks.sh\n", "time ./stacks.sh" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## aftrRAD empirical analysis\n", "* AftrRAD.pl - 20164m43s\n", "* Genotype.pl - 879m18s\n", "* FilterSNPs.pl - 574m31s" ] }, { "cell_type": "code", "execution_count": 14, "metadata": { "collapsed": false, "scrolled": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl AftrRAD.pl minDepth-6 P2-noP2 minIden-85 re-TGCAG stringLength-99 dplexedData-1 maxProcesses-40\n", "local::lib is up to date. (2.000019)\n", "Formatting data file 30556_thamno_R1_.fastq\n", "Formatting data file 32082_przewalskii_R1_.fastq\n", "Formatting data file 29154_superba_R1_.fastq\n", "Formatting data file 40578_rex_R1_.fastq\n", "Formatting data file 33588_przewalskii_R1_.fastq\n", "Formatting data file 33413_thamno_R1_.fastq\n", "Formatting data file 41478_cyathophylloides_R1_.fastq\n", "Formatting data file 35855_rex_R1_.fastq\n", "Formatting data file 39618_rex_R1_.fastq\n", "Formatting data file 38362_rex_R1_.fastq\n", "Formatting data file 35236_rex_R1_.fastq\n", "Formatting data file 41954_cyathophylloides_R1_.fastq\n", "Formatting data file 30686_cyathophylla_R1_.fastq\n", "\n", "\n", "Arguments entered are...\n", "minParalog\t5\n", "minDepth\t6\n", "MaxH\t90\n", "P2\tnoP2\n", "BarcodePath\tBarcodes/\n", "minQual\t20\n", "re\tTGCAG\n", "numIndels\t3\n", "Phred\t33\n", "minIden\t85\n", "maxProcesses\t40\n", "stringLength\t99\n", "dplexedData\t1\n", "Help\t0\n", "DataPath\tData/\n", "\n", "\n", "Running AftrRAD...\n", "\n", "Recognized 13 data files.\n", "\n", "File names are...\n", " 30556_thamno_R1_.fastq.txt 32082_przewalskii_R1_.fastq.txt 29154_superba_R1_.fastq.txt 40578_rex_R1_.fastq.txt 33588_przewalskii_R1_.fastq.txt 33413_thamno_R1_.fastq.txt 41478_cyathophylloides_R1_.fastq.txt 35855_rex_R1_.fastq.txt 39618_rex_R1_.fastq.txt 38362_rex_R1_.fastq.txt 35236_rex_R1_.fastq.txt 41954_cyathophylloides_R1_.fastq.txt 30686_cyathophylla_R1_.fastq.txt \n", "\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 5.0 (file 33413_thamno_R1_.fastq.txt) are...\n", " CAGGAC \n", "\n", "The names you entered for data file 5.0 (file 33413_thamno_R1_.fastq.txt) are...\n", " 33413_thamno_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 33413_thamno_R1_.fastq.txt.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 2.0 (file 29154_superba_R1_.fastq.txt) are...\n", " CAGAAA \n", "\n", "The names you entered for data file 2.0 (file 29154_superba_R1_.fastq.txt) are...\n", " 29154_superba_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 29154_superba_R1_.fastq.txt.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "\n", "Identifying unique sequences for each individual.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 8.0 (file 39618_rex_R1_.fastq.txt) are...\n", " CTATCA \n", "\n", "The names you entered for data file 8.0 (file 39618_rex_R1_.fastq.txt) are...\n", " 39618_rex_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 39618_rex_R1_.fastq.txt.\n", "170622 TempFiles/SubTemp_5/UniqueWithCountsIndividual33413_thamno_R1_.txt\n", "\n", "Identifying unique sequences for each individual.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 1.0 (file 32082_przewalskii_R1_.fastq.txt) are...\n", " TTCCAG \n", "\n", "The names you entered for data file 1.0 (file 32082_przewalskii_R1_.fastq.txt) are...\n", " 32082_przewalskii_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 32082_przewalskii_R1_.fastq.txt.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "146010 TempFiles/SubTemp_2/UniqueWithCountsIndividual29154_superba_R1_.txt\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 4.0 (file 33588_przewalskii_R1_.fastq.txt) are...\n", " CGAGCG \n", "\n", "The names you entered for data file 4.0 (file 33588_przewalskii_R1_.fastq.txt) are...\n", " 33588_przewalskii_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 33588_przewalskii_R1_.fastq.txt.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "\n", "Identifying unique sequences for each individual.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 12.0 (file 30686_cyathophylla_R1_.fastq.txt) are...\n", " CGCCTG \n", "\n", "The names you entered for data file 12.0 (file 30686_cyathophylla_R1_.fastq.txt) are...\n", " 30686_cyathophylla_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 30686_cyathophylla_R1_.fastq.txt.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "\n", "Identifying unique sequences for each individual.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "172534 TempFiles/SubTemp_8/UniqueWithCountsIndividual39618_rex_R1_.txt\n", "\n", "Identifying unique sequences for each individual.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1072.\n", "Use of uninitialized value $TotalBarcodeNonMatches in concatenation (.) or string at AftrRAD.pl line 1073.\n", "\n", "Identifying unique sequences for each individual.\n", "175494 TempFiles/SubTemp_1/UniqueWithCountsIndividual32082_przewalskii_R1_.txt\n", "190036 TempFiles/SubTemp_4/UniqueWithCountsIndividual33588_przewalskii_R1_.txt\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 9.0 (file 38362_rex_R1_.fastq.txt) are...\n", " CCCACA \n", "\n", "The names you entered for data file 9.0 (file 38362_rex_R1_.fastq.txt) are...\n", " 38362_rex_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 38362_rex_R1_.fastq.txt.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 7.0 (file 35855_rex_R1_.fastq.txt) are...\n", " ATAACT \n", "\n", "The names you entered for data file 7.0 (file 35855_rex_R1_.fastq.txt) are...\n", " 35855_rex_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 35855_rex_R1_.fastq.txt.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 0.0 (file 30556_thamno_R1_.fastq.txt) are...\n", " CCGTAT \n", "\n", "The names you entered for data file 0.0 (file 30556_thamno_R1_.fastq.txt) are...\n", " 30556_thamno_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 30556_thamno_R1_.fastq.txt.\n", "241443 TempFiles/SubTemp_12/UniqueWithCountsIndividual30686_cyathophylla_R1_.txt\n", "\n", "Identifying unique sequences for each individual.\n", "\n", "Identifying unique sequences for each individual.\n", "\n", "Identifying unique sequences for each individual.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 3.0 (file 40578_rex_R1_.fastq.txt) are...\n", " TATCTA \n", "\n", "The names you entered for data file 3.0 (file 40578_rex_R1_.fastq.txt) are...\n", " 40578_rex_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 40578_rex_R1_.fastq.txt.\n", "166314 TempFiles/SubTemp_9/UniqueWithCountsIndividual38362_rex_R1_.txt\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 10.0 (file 35236_rex_R1_.fastq.txt) are...\n", " GTCTTT \n", "\n", "The names you entered for data file 10.0 (file 35236_rex_R1_.fastq.txt) are...\n", " 35236_rex_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 35236_rex_R1_.fastq.txt.\n", "\n", "Identifying unique sequences for each individual.\n", "266368 TempFiles/SubTemp_7/UniqueWithCountsIndividual35855_rex_R1_.txt\n", "274366 TempFiles/SubTemp_0/UniqueWithCountsIndividual30556_thamno_R1_.txt\n", "\n", "Identifying unique sequences for each individual.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 11.0 (file 41954_cyathophylloides_R1_.fastq.txt) are...\n", " GCACCT \n", "\n", "The names you entered for data file 11.0 (file 41954_cyathophylloides_R1_.fastq.txt) are...\n", " 41954_cyathophylloides_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 41954_cyathophylloides_R1_.fastq.txt.\n", "\n", "Warning...only one line recognized in barcode file 30686_cyathophylla_R1_.fastq.txt. Check format of this file if it contains more than one sample.\n", "The barcodes you entered for data file 6.0 (file 41478_cyathophylloides_R1_.fastq.txt) are...\n", " AGGCGA \n", "\n", "The names you entered for data file 6.0 (file 41478_cyathophylloides_R1_.fastq.txt) are...\n", " 41478_cyathophylloides_R1_ \n", "\n", "\n", "Demultiplexing samples for data file 41478_cyathophylloides_R1_.fastq.txt.\n", "253097 TempFiles/SubTemp_3/UniqueWithCountsIndividual40578_rex_R1_.txt\n", "417767 TempFiles/SubTemp_10/UniqueWithCountsIndividual35236_rex_R1_.txt\n", "\n", "Identifying unique sequences for each individual.\n", "\n", "Identifying unique sequences for each individual.\n", "318528 TempFiles/SubTemp_11/UniqueWithCountsIndividual41954_cyathophylloides_R1_.txt\n", "205456 TempFiles/SubTemp_6/UniqueWithCountsIndividual41478_cyathophylloides_R1_.txt\n", "\n", "Identifying all unique sequences within the dataset.\n", "\n", "Creating file to test mean read counts.\n", "Testing nonzero means in read counts to identify error reads.\n", "\n", "Performing heuristic search of all pairwise comparisons to identify potentially allelic pairs.\n", "Rapidly searching unique sequence number 1000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 2000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 3000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 4000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 5000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 6000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 7000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 8000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 9000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 10000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 11000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 12000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 13000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 14000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 15000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 16000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 17000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 18000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 19000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 20000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 21000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 22000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 23000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 24000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 25000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 26000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 27000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 28000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 29000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 30000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 31000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 32000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 33000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 34000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 35000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 36000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 37000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 38000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 39000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 40000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 41000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 42000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 43000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 44000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 45000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 46000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 47000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 48000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 49000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 50000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 51000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 52000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 53000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 54000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 55000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 56000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 57000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 58000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 59000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 60000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 61000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 62000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 63000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 64000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 65000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 66000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 67000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 68000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 69000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 70000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 71000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 72000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 73000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 74000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 75000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 76000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 77000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 78000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 79000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 80000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 81000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 82000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 83000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 84000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 85000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 86000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 87000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 88000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 89000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 90000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 91000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 92000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 93000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 94000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 95000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 96000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 97000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 98000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 99000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 100000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 101000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 102000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 103000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 104000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 105000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 106000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 107000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 108000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 109000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 110000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 111000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 112000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 113000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 114000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 115000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 116000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 117000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 118000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 119000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 120000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 121000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 122000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 123000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 124000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 125000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 126000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 127000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 128000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 129000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 130000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 131000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 132000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 133000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 134000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 135000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 136000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 137000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 138000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 139000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 140000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 141000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 142000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 143000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 144000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 145000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 146000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 147000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 148000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 149000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 150000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 151000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 152000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 153000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 154000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 155000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 156000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 157000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 158000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 159000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 160000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 161000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 162000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 163000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 164000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 165000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 166000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 167000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 168000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 169000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 170000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 171000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 172000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 173000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 174000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 175000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 176000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 177000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 178000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 179000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 180000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 181000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 182000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 183000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 184000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 185000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 186000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 187000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 188000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 189000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 190000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 191000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 192000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 193000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 194000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 195000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 196000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 197000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 198000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 199000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 200000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 201000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 202000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 203000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 204000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 205000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 206000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 207000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 208000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 209000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 210000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 211000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 212000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 213000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 214000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 215000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 216000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 217000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 218000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 219000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 220000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 221000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 222000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 223000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 224000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 225000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 226000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 227000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 228000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 229000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 230000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 231000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 232000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 233000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 234000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 235000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 236000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 237000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 238000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 239000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 240000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 241000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 242000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 243000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 244000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 245000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 246000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 247000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 248000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 249000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 250000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 251000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 252000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 253000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 254000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 255000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 256000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 257000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 258000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 259000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 260000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 261000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 262000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 263000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 264000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 265000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 266000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 267000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 268000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 269000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 270000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 271000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 272000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 273000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 274000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 275000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 276000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 277000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 278000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 279000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 280000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 281000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 282000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 283000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 284000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 285000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 286000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 287000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 288000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 289000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 290000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 291000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 292000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 293000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 294000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 295000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 296000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 297000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 298000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 299000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 300000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 301000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 302000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 303000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 304000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 305000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 306000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 307000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 308000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 309000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 310000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 311000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 312000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 313000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 314000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 315000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 316000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 317000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 318000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 319000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 320000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 321000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 322000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 323000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 324000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 325000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 326000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 327000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 328000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 329000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 330000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 331000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 332000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 333000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 334000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 335000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 336000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 337000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 338000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 339000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 340000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 341000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 342000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 343000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 344000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 345000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 346000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 347000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 348000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 349000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 350000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 351000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 352000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 353000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 354000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 355000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 356000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 357000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 358000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 359000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 360000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 361000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 362000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 363000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 364000 of 365732 for potentially allelic read pairs.\n", "Rapidly searching unique sequence number 365000 of 365732 for potentially allelic read pairs.\n", "\n", "Number of pairwise comparisons to align and evaluate is 3712847\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 40000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 80000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 120000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 160000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 200000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 240000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 280000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 320000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 360000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 400000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 440000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 480000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 520000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 560000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 600000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 640000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 680000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 720000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 760000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 800000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 840000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 880000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 920000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 960000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1000000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1040000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1080000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1120000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1160000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1200000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1240000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1280000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1320000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1360000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1400000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1440000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1480000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1520000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1560000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1600000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1640000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1680000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1720000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1760000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1800000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1840000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1880000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1920000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 1960000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2000000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2040000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2080000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2120000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2160000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2200000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2240000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2280000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2320000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2360000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2400000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2440000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2480000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2520000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2560000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2600000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2640000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2680000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2720000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2760000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2800000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2840000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2880000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2920000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 2960000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3000000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3040000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3080000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3120000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3160000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3200000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3240000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3280000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3320000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3360000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3400000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3440000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3480000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3520000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3560000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3600000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3640000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "Working on alignment 3680000 of 3712847.\n", "\n", "Scoring each pairwise alignment. Alignments exceeding 85 % similarity will be retained.\n", "Parsing aligned sequences into candidate loci.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 4000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 8000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 12000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 16000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 20000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 24000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 28000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 32000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 36000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 40000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 44000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 48000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 52000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 56000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Working on alignment 60000 of 61742.\n", "Finished writing the mafft output.\n", "Identifying and removing paralogous loci.\n", "\n", "\n", "Loci have been identified. Aligned polymorphic loci are stored in the Output directory. Read counts for the alleles at these loci can be found for each individual in the files stored in TempFiles/RawReadCountFiles.\n", "\n", "Demultiplexing and other general information can be found for each dataset analyzed in the Output/RunInfo directory.\n", "\n", "Next, run Genotype.pl, followed by FilterSNPs.pl to genotype all individuals and output SNPs for subsequent analyses.\n", "\n", "\n", "real\t20164m43.822s\n", "user\t2634m47.374s\n", "sys\t620257m19.431s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl Genotype.pl MinReads-6 subset-0 maxProcesses-40\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "subsetfile\tSamplesForSubset.txt\n", "pvalLow\t0.0001\n", "pvalThresh\t100\n", "subset\t0\n", "maxProcesses\t40\n", "MinReads\t6\n", "pvalHigh\t1e-05\n", "Help\t0\n", "\n", "\n", "Running Genotype.pl...\n", "\n", "Scoring genotypes at all nonparalogous loci.\n", "\n", "Currently genotyping sample...39618_rex_R1_\t35855_rex_R1_\t41954_cyathophylloides_R1_\t33413_thamno_R1_\t38362_rex_R1_\t29154_superba_R1_\t30556_thamno_R1_\t33588_przewalskii_R1_\t40578_rex_R1_\t32082_przewalskii_R1_\t30686_cyathophylla_R1_\t35236_rex_R1_\t41478_cyathophylloides_R1_\tRunning R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Running R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "Finished R\n", "cat: TempFiles/SubGenotypes_13/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_14/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_15/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_16/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_17/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_18/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_19/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_20/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_21/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_22/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_23/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_24/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_25/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_26/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_27/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_28/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_29/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_30/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_31/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_32/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_33/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_34/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_35/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_36/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_37/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_38/SubGenotypes.txt: No such file or directory\n", "cat: TempFiles/SubGenotypes_39/SubGenotypes.txt: No such file or directory\n", "Creating SNPMatrixAll from R Script\n", "\n", "\n", "Checking for individuals with large amounts of missing data (have >2 StDev more missing data than the average)\n", "Identifying samples with >2 stdev missing data\n", "Read 13 items\n", "Read 13 items\n", "null device \n", " 1 \n", "No samples were flagged as potentially bad samples. Would like to remove any samples from further analyses anyway? (y/n)\n", "All individuals will be analyzed.\n", "All samples were included in analyses.\n", "Reading SNPMatrixAll file\n", "Identifying monomorphic loci\n", "107475 TempFiles/MonomorphicsNoParalogs.txt\n", "Updating Genotypes.txt file\n", "\n", "Identifying SNPs and determining the position of each SNP along the read.\n", "Plotting SNP locations\n", "Read 242297 items\n", "\n", "Loci have all been genotyped, and scored genotypes are stored in the file 'Genotypes.txt' in the TempFiles folder.\n", "\n", "The locations of each SNP are plotted in the file SNPLocations.pdf in the Output/RunInfo folder. Use this plot to choose a cut-off value for omitting SNPs due to artifacts toward the ends of reads.\n", "\n", "Next, run 'FilterSNPs.pl' to filter and output SNPs and monomorphic loci based on 1.) the proportion of samples genotyped at each locus and 2.) SNP positions along the reads.\n", "\n", "real\t879m18.452s\n", "user\t880m50.057s\n", "sys\t6m16.364s\n", "\n", "export PATH=/home/iovercast/manuscript-analysis/perl5/bin:$PATH; cpanm --local-lib=/home/iovercast/manuscript-analysis//perl5 local::lib && eval $(perl -I /home/iovercast/manuscript-analysis//perl5/lib/perl5/ -Mlocal::lib); time perl FilterSNPs.pl pctScored-17\n", "local::lib is up to date. (2.000019)\n", "\n", "\n", "Arguments entered are...\n", "maxSNP\t0\n", "MinReads\t6\n", "pctScored\t17\n", "Help\t0\n", "\n", "\n", "Running FilterSNPs.pl...\n", "\n", "Identifying all SNPs that are scored in 17% of the samples\n", "Individual SNPs, haplotypes, and monomorphic loci have been printed to files located in Output/Genotypes.\n", "These data can be further formatted using the scripts in the 'Formatting' directory\n", "\n", "real\t574m31.944s\n", "user\t574m40.141s\n", "sys\t0m7.166s\n", "\n" ] } ], "source": [ "import subprocess\n", "import gzip\n", "\n", "## Set up directory structures. change the force flag if you want to\n", "## blow everything away and restart\n", "# force = True\n", "force = \"\"\n", "\n", "if force and os.path.exists(AFTRRAD_OUTPUT):\n", " shutil.rmtree(AFTRRAD_OUTPUT)\n", "if not os.path.exists(AFTRRAD_OUTPUT):\n", " os.makedirs(AFTRRAD_OUTPUT)\n", "os.chdir(AFTRRAD_OUTPUT)\n", "\n", "## Prep empirical data by copying it to a local folder called DemultiplexedFiles\n", "## in this way you don't need the Barcodes/Data dir\n", "demux_dir = os.path.join(AFTRRAD_OUTPUT, \"DemultiplexedFiles/\")\n", "if force and os.path.exists(demux_dir):\n", " shutil.rmtree(demux_dir)\n", "if not os.path.exists(demux_dir):\n", " os.makedirs(demux_dir)\n", "\n", "IPYRAD_EDITS_DIR = os.path.join(IPYRAD_OUTPUT, \"REALDATA_edits/\")\n", "infiles = [ff for ff in glob.glob(IPYRAD_EDITS_DIR+\"*_R1_*\")]\n", "## Don't recopy the data over and over again while testing\n", "if not os.path.exists(demux_dir+infiles[0].split(\"/\")[-1]):\n", " for f in infiles:\n", " shutil.copy2(f, demux_dir+f.split(\"/\")[-1])\n", "\n", "## AfterRAD is incredibly picky about where scripts and binaries are so you\n", "## have to do a bunch of annoying housekeeping.\n", "cmd = \"cp -rf {}/* {}\".format(os.path.join(AFTRRAD_DIR, \"AftrRAD/AftrRADv5.0\"), AFTRRAD_OUTPUT)\n", "os.system(cmd)\n", "shutil.copy2(os.path.join(AFTRRAD_DIR, \"dnaMatrix\"), AFTRRAD_OUTPUT)\n", "shutil.copy2(os.path.join(AFTRRAD_DIR, \"ACANA\"), AFTRRAD_OUTPUT)\n", "\n", "## This first line of nonsense is to make it so perl can find the Parallel::ForkManager library\n", "base = \"export PATH={}perl5/bin:$PATH; cpanm --local-lib={}/perl5 local::lib && eval $(perl -I {}/perl5/lib/perl5/ -Mlocal::lib); \".format(WORK_DIR, WORK_DIR, WORK_DIR)\n", "\n", "## Good sub for testing\n", "#cmd = base + \"time perl AftrRAD.pl -h\"\n", "#output = subprocess.check_output(cmd, shell=True, stderr=subprocess.STDOUT)\n", "#print(output)\n", "\n", "## numIndels-3 What do here?\n", "## stringLength-99 We don't care about homoplymers here, so set it very high\n", "## maxH-90 ??\n", "## AftrRAD manu analysis settings - P2: noP2; minDepth: 5; numIndel: 3; MinReads: 5\n", "cmd = base + \"time perl AftrRAD.pl minDepth-6 P2-noP2 minIden-85 \" \\\n", " + \"re-TGCAG stringLength-99 dplexedData-1 maxProcesses-40\"\n", "print(cmd)\n", "output = \"\"\n", "output = subprocess.check_output(cmd, shell=True, stderr=subprocess.STDOUT)\n", "print(output)\n", "\n", "## MinReads is mindepth_statistical\n", "cmd = base + \"time perl Genotype.pl MinReads-6 subset-0 maxProcesses-40\"\n", "print(cmd)\n", "output = subprocess.check_output(cmd, shell=True, stderr=subprocess.STDOUT)\n", "print(output)\n", "\n", "## pctScored is min_samples_locus as a percent of total samples, here since\n", "## we use 2 for min_samples_locus and there are 12 simulated samples we use 17%\n", "cmd = base + \"time perl FilterSNPs.pl pctScored-17\"\n", "print(cmd)\n", "output = subprocess.check_output(cmd, shell=True, stderr=subprocess.STDOUT)\n", "\n", "print(output)" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "### Post-process aftrRAD output so we actually can get a vcf file" ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": true }, "outputs": [], "source": [ "%%bash -s \"$WORK_DIR\" -s \"$AFTRRAD_DIR\"\n", "export PATH=\"$1/miniconda/bin:$PATH\"; export \"WORK_DIR=$1\"; export \"AFTRRAD_DIR=$2\"\n", "cd $AFTRRAD_DIR/REALDATA/Formatting\n", "\n", "perl OutputFasta.pl SNPsOnly-1\n", "snp-sites -v -o REALDATA.vcf SNPMatrix_*.All.fasta" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "## dDocent empirical analysis\n", "Runtime: ~ 15 minutes" ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": false, "scrolled": true }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "Doing - no\n", " New Assembly: simno\n", "ipyrad -p params-simno.txt -s 1\n", "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.4.0]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: simno\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Demultiplexing fastq data to Samples\n", " [####################] 100% chunking large files | 0:00:00 \n", " [####################] 100% sorting reads | 0:00:44 \n", " [####################] 100% writing/compressing | 0:00:02 \n", "\n", "\n", "\n", "real\t0m56.558s\n", "user\t0m1.724s\n", "sys\t0m0.437s\n", "Renaming ./1A_0_R1_.fastq.gz -> Pop1_SampleA.F.fq.gz\n", "Renaming ./3J_0_R1_.fastq.gz -> Pop3_SampleJ.F.fq.gz\n", "Renaming ./1B_0_R1_.fastq.gz -> Pop1_SampleB.F.fq.gz\n", "Renaming ./3K_0_R1_.fastq.gz -> Pop3_SampleK.F.fq.gz\n", "Renaming ./2F_0_R1_.fastq.gz -> Pop2_SampleF.F.fq.gz\n", "Renaming ./1D_0_R1_.fastq.gz -> Pop1_SampleD.F.fq.gz\n", "Renaming ./1C_0_R1_.fastq.gz -> Pop1_SampleC.F.fq.gz\n", "Renaming ./3L_0_R1_.fastq.gz -> Pop3_SampleL.F.fq.gz\n", "Renaming ./2E_0_R1_.fastq.gz -> Pop2_SampleE.F.fq.gz\n", "Renaming ./3I_0_R1_.fastq.gz -> Pop3_SampleI.F.fq.gz\n", "Renaming ./2G_0_R1_.fastq.gz -> Pop2_SampleG.F.fq.gz\n", "Renaming ./2H_0_R1_.fastq.gz -> Pop2_SampleH.F.fq.gz\n", "export LD_LIBRARY_PATH=/home/iovercast/manuscript-analysis/dDocent//freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; export PATH=/home/iovercast/manuscript-analysis/dDocent/:$PATH; dDocent simno-config.txt\n", "dDocent 2.24 \n", "\n", "Contact jpuritz@gmail.com with any problems \n", "\n", " \n", "Checking for required software\n", "\n", "All required software is installed!\n", "dDocent will require input during the assembly stage. Please wait until prompt says it is safe to move program to the background.\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "Removing the _1 character and replacing with /1 in the name of every sequence\n", "Trimming reads and simultaneously assembling reference sequences\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "sort: write failed: standard output: Broken pipe\n", "sort: write error\n", "\f", " \n", " Number of Unique Sequences with More than X Coverage (Counted within individuals) \n", " \n", " 138000 +-+---------************+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " + + ***********************************+ + + + + \n", " 136000 +-+ ********* +-+ \n", " | *** | \n", " | ***** | \n", " 134000 +-+ *** +-+ \n", " | *** | \n", " 132000 +-+ * +-+ \n", " | *** | \n", " 130000 +-+ ** +-+ \n", " | ** | \n", " 128000 +-+ ** +-+ \n", " | ** | \n", " | * | \n", " 126000 +-+ *** +-+ \n", " | * | \n", " 124000 +-+ ** +-+ \n", " | * | \n", " 122000 +-+ ** +-+ \n", " | * | \n", " | **| \n", " 120000 +-+ +-+ \n", " + + + + + + + + + * \n", " 118000 +-+---------+-----------+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " 1 2 3 4 5 6 7 8 9 10 \n", " Coverage \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. In essence, you are picking a minimum (within individual) coverage level for a read (allele) to be used in the reference assembly\n", "\f", " \n", " Number of Unique Sequences present in more than X Individuals \n", " \n", " 20000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " + + + + + + + + + \n", " ****** | \n", " | *** | \n", " 18000 +-+ *** +-+ \n", " | **** | \n", " | *** | \n", " | *** | \n", " 16000 +-+ **** +-+ \n", " | * | \n", " | ******* | \n", " 14000 +-+ ***** +-+ \n", " | **** | \n", " | ***** | \n", " | **** | \n", " 12000 +-+ ** +-+ \n", " | ********* | \n", " | ***** | \n", " | ***** | \n", " 10000 +-+ ****** +-+ \n", " | ** | \n", " | ***************************| \n", " + + + + + + + + * \n", " 8000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " 2 2.5 3 3.5 4 4.5 5 5.5 6 \n", " Number of Individuals \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. Pick point right before the assymptote. A good starting cutoff might be 10% of the total number of individuals\n", "At this point, all configuration information has been entered and dDocent may take several hours to run.\n", "It is recommended that you move this script to a background operation and disable terminal input and output.\n", "All data and logfiles will still be recorded.\n", "To do this:\n", "Press control and Z simultaneously\n", "Type 'bg' without the quotes and press enter\n", "Type 'disown -h' again without the quotes and press enter\n", "\n", "Now sit back, relax, and wait for your analysis to finish.\n", "TrimmomaticSE: Started with arguments: -threads 40 -phred33 uniq.fq uniq.fq1 ILLUMINACLIP:/home/iovercast/manuscript-analysis/dDocent/TruSeq2-PE.fa:2:30:10 MINLEN:68\n", "Using PrefixPair: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT' and 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTAATGATACGGCGACCACCGAGATCTACAC'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTCAAGCAGAAGACGGCATACGA'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGCCGTATCATT'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGGTTCAGCAGGAATGCCGAGACCGATCTCGTATGCCGTCTTCTGCTTG'\n", "Using Long Clipping Sequence: 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT'\n", "ILLUMINACLIP: Using 1 prefix pairs, 6 forward/reverse sequences, 0 forward only sequences, 0 reverse only sequences\n", "Input Reads: 18850 Surviving: 18850 (100.00%) Dropped: 0 (0.00%)\n", "TrimmomaticSE: Completed successfully\n", "================================================================\n", "Program: CD-HIT, V4.6 (+OpenMP), Sep 21 2016, 20:52:31\n", "Command: cd-hit-est -i totalover.fasta -o\n", " reference.fasta.original -M 0 -T 0 -c 0.85\n", "\n", "Started: Sat Oct 1 18:01:38 2016\n", "================================================================\n", " Output \n", "----------------------------------------------------------------\n", "total number of CPUs in the system is 40\n", "Actual number of CPUs to be used: 40\n", "\n", "total seq: 18850\n", "longest and shortest : 91 and 91\n", "Total letters: 1715350\n", "Sequences have been sorted\n", "\n", "Approximated minimal memory consumption:\n", "Sequence : 4M\n", "Buffer : 40 X 12M = 491M\n", "Table : 2 X 17M = 34M\n", "Miscellaneous : 4M\n", "Total : 534M\n", "\n", "Table limit with the given memory limit:\n", "Max number of representatives: 366227\n", "Max number of word counting entries: 8747853\n", "\n", "# comparing sequences from 0 to 448\n", "---------- new table with 444 representatives\n", "# comparing sequences from 448 to 886\n", "---------- 333 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 553 to 988\n", "---------- 256 remaining sequences to the next cycle\n", "---------- new table with 171 representatives\n", "# comparing sequences from 732 to 1163\n", "---------- 189 remaining sequences to the next cycle\n", "---------- new table with 229 representatives\n", "# comparing sequences from 974 to 1399\n", "---------- 157 remaining sequences to the next cycle\n", "---------- new table with 253 representatives\n", "# comparing sequences from 1242 to 1661\n", "---------- 110 remaining sequences to the next cycle\n", "---------- new table with 282 representatives\n", "# comparing sequences from 1551 to 1962\n", "---------- 87 remaining sequences to the next cycle\n", "---------- new table with 286 representatives\n", "# comparing sequences from 1875 to 2279\n", "---------- 134 remaining sequences to the next cycle\n", "---------- new table with 239 representatives\n", "# comparing sequences from 2145 to 2542\n", "---------- 83 remaining sequences to the next cycle\n", "---------- new table with 268 representatives\n", "# comparing sequences from 2459 to 2849\n", "---------- 274 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 2575 to 2962\n", "---------- 201 remaining sequences to the next cycle\n", "---------- new table with 152 representatives\n", "# comparing sequences from 2761 to 3144\n", "---------- 260 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 2884 to 3264\n", "---------- 95 remaining sequences to the next cycle\n", "---------- new table with 232 representatives\n", "# comparing sequences from 3169 to 3542\n", "---------- 120 remaining sequences to the next cycle\n", "---------- new table with 204 representatives\n", "# comparing sequences from 3422 to 3789\n", "---------- 60 remaining sequences to the next cycle\n", "---------- new table with 256 representatives\n", "# comparing sequences from 3729 to 4089\n", "---------- 93 remaining sequences to the next cycle\n", "---------- new table with 206 representatives\n", "# comparing sequences from 3996 to 4349\n", "---------- 212 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 4137 to 4487\n", "99.8%---------- new table with 255 representatives\n", "# comparing sequences from 4487 to 4828\n", "---------- 23 remaining sequences to the next cycle\n", "---------- new table with 235 representatives\n", "# comparing sequences from 4805 to 5139\n", "---------- 111 remaining sequences to the next cycle\n", "---------- new table with 159 representatives\n", "# comparing sequences from 5028 to 5357\n", "---------- 115 remaining sequences to the next cycle\n", "---------- new table with 148 representatives\n", "# comparing sequences from 5242 to 5566\n", "---------- 165 remaining sequences to the next cycle\n", "---------- new table with 110 representatives\n", "# comparing sequences from 5401 to 5721\n", "---------- 164 remaining sequences to the next cycle\n", "---------- new table with 116 representatives\n", "# comparing sequences from 5557 to 5873\n", "---------- 141 remaining sequences to the next cycle\n", "---------- new table with 123 representatives\n", "# comparing sequences from 5732 to 6044\n", "---------- 163 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5881 to 6189\n", "---------- 141 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6048 to 6352\n", "---------- 130 remaining sequences to the next cycle\n", "---------- new table with 110 representatives\n", "# comparing sequences from 6222 to 6522\n", "---------- 79 remaining sequences to the next cycle\n", "---------- new table with 136 representatives\n", "# comparing sequences from 6443 to 6738\n", "---------- 120 remaining sequences to the next cycle\n", "---------- new table with 108 representatives\n", "# comparing sequences from 6618 to 6909\n", "---------- 107 remaining sequences to the next cycle\n", "---------- new table with 113 representatives\n", "# comparing sequences from 6802 to 7088\n", "---------- 110 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6978 to 7260\n", "---------- 107 remaining sequences to the next cycle\n", "---------- new table with 111 representatives\n", "# comparing sequences from 7153 to 7431\n", "---------- 97 remaining sequences to the next cycle\n", "---------- new table with 106 representatives\n", "# comparing sequences from 7334 to 7608\n", "---------- 75 remaining sequences to the next cycle\n", "---------- new table with 115 representatives\n", "# comparing sequences from 7533 to 7802\n", "---------- 110 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7692 to 7957\n", "---------- 86 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7871 to 8132\n", "---------- 86 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8046 to 8303\n", "---------- 66 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8237 to 8489\n", "---------- 59 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8430 to 8678\n", "---------- 34 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8644 to 8887\n", "---------- 48 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8839 to 9077\n", "---------- 34 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9043 to 9276\n", "---------- 43 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9233 to 9461\n", "---------- 39 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9422 to 9646\n", "---------- 28 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9618 to 9837\n", "---------- 16 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9821 to 10035\n", "..........---------- new table with 93 representatives\n", "# comparing sequences from 10035 to 10244\n", "........---------- new table with 100 representatives\n", "# comparing sequences from 10244 to 10448\n", "---------- 11 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 10437 to 10637\n", "...........---------- new table with 100 representatives\n", "# comparing sequences from 10637 to 10832\n", ".....---------- new table with 99 representatives\n", "# comparing sequences from 10832 to 11022\n", "........---------- new table with 77 representatives\n", "# comparing sequences from 11022 to 11208\n", ".........---------- new table with 78 representatives\n", "# comparing sequences from 11208 to 11389\n", "............---------- new table with 84 representatives\n", "# comparing sequences from 11389 to 11566\n", "...........---------- new table with 70 representatives\n", "# comparing sequences from 11566 to 11739\n", "........---------- new table with 85 representatives\n", "# comparing sequences from 11739 to 11908\n", "..........---------- new table with 71 representatives\n", "# comparing sequences from 11908 to 12073\n", "........---------- new table with 70 representatives\n", "# comparing sequences from 12073 to 12234\n", "...---------- new table with 48 representatives\n", "# comparing sequences from 12234 to 12391\n", "..........---------- new table with 70 representatives\n", "# comparing sequences from 12391 to 12544\n", ".........---------- new table with 68 representatives\n", "# comparing sequences from 12544 to 12694\n", "......---------- new table with 51 representatives\n", "# comparing sequences from 12694 to 12840\n", "...---------- new table with 46 representatives\n", "# comparing sequences from 12840 to 12983\n", "........---------- new table with 59 representatives\n", "# comparing sequences from 12983 to 13122\n", ".......---------- new table with 48 representatives\n", "# comparing sequences from 13122 to 13258\n", ".......---------- new table with 49 representatives\n", "# comparing sequences from 13258 to 13391\n", "......---------- new table with 37 representatives\n", "# comparing sequences from 13391 to 13520\n", ".......---------- new table with 41 representatives\n", "# comparing sequences from 13520 to 13646\n", ".......---------- new table with 40 representatives\n", "# comparing sequences from 13646 to 13769\n", "........---------- new table with 43 representatives\n", "# comparing sequences from 13769 to 13889\n", ".......---------- new table with 31 representatives\n", "# comparing sequences from 13889 to 14007\n", ".......---------- new table with 34 representatives\n", "# comparing sequences from 14007 to 14122\n", "...........---------- new table with 34 representatives\n", "# comparing sequences from 14122 to 14234\n", ".........---------- new table with 39 representatives\n", "# comparing sequences from 14234 to 14343\n", ".......---------- new table with 37 representatives\n", "# comparing sequences from 14343 to 14450\n", ".....---------- new table with 39 representatives\n", "# comparing sequences from 14450 to 14554\n", "......---------- new table with 35 representatives\n", "# comparing sequences from 14554 to 14656\n", ".......---------- new table with 30 representatives\n", "# comparing sequences from 14656 to 14755\n", ".......---------- new table with 29 representatives\n", "# comparing sequences from 14755 to 14852\n", ".....---------- new table with 29 representatives\n", "# comparing sequences from 14852 to 14947\n", "........---------- new table with 34 representatives\n", "# comparing sequences from 14947 to 15039\n", ".......---------- new table with 22 representatives\n", "# comparing sequences from 15039 to 15129\n", "....---------- new table with 28 representatives\n", "# comparing sequences from 15129 to 15217\n", "......---------- new table with 20 representatives\n", "# comparing sequences from 15217 to 15303\n", "....---------- new table with 30 representatives\n", "# comparing sequences from 15303 to 15387\n", ".......---------- new table with 27 representatives\n", "# comparing sequences from 15387 to 15469\n", ".......---------- new table with 23 representatives\n", "# comparing sequences from 15469 to 15549\n", "..---------- new table with 22 representatives\n", "# comparing sequences from 15549 to 15627\n", ".....---------- new table with 18 representatives\n", "# comparing sequences from 15627 to 15703\n", ".......---------- new table with 26 representatives\n", "# comparing sequences from 15703 to 15777\n", "....---------- new table with 22 representatives\n", "# comparing sequences from 15777 to 15850\n", "...---------- new table with 18 representatives\n", "# comparing sequences from 15850 to 15921\n", "......---------- new table with 19 representatives\n", "# comparing sequences from 15921 to 15990\n", "......---------- new table with 17 representatives\n", "# comparing sequences from 15990 to 16058\n", ".---------- new table with 13 representatives\n", "# comparing sequences from 16058 to 16124\n", "....---------- new table with 19 representatives\n", "# comparing sequences from 16124 to 16188\n", "..---------- new table with 18 representatives\n", "# comparing sequences from 16188 to 16251\n", "...---------- new table with 18 representatives\n", "# comparing sequences from 16251 to 16312\n", "....---------- new table with 16 representatives\n", "# comparing sequences from 16312 to 16372\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 16372 to 16431\n", "......---------- new table with 21 representatives\n", "# comparing sequences from 16431 to 16488\n", "...---------- new table with 11 representatives\n", "# comparing sequences from 16488 to 16544\n", "...---------- new table with 12 representatives\n", "# comparing sequences from 16544 to 16598\n", "..---------- new table with 10 representatives\n", "# comparing sequences from 16598 to 16651\n", "..---------- new table with 9 representatives\n", "# comparing sequences from 16651 to 16703\n", ".......---------- new table with 16 representatives\n", "# comparing sequences from 16703 to 16754\n", "....---------- new table with 15 representatives\n", "# comparing sequences from 16754 to 16803\n", ".....---------- new table with 12 representatives\n", "# comparing sequences from 16803 to 16851\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 16851 to 16898\n", ".---------- new table with 7 representatives\n", "# comparing sequences from 16898 to 16944\n", "...---------- new table with 8 representatives\n", "# comparing sequences from 16944 to 16989\n", "..---------- new table with 10 representatives\n", "# comparing sequences from 16989 to 17033\n", "...---------- new table with 10 representatives\n", "# comparing sequences from 17033 to 17076\n", "...---------- new table with 13 representatives\n", "# comparing sequences from 17076 to 17118\n", "...---------- new table with 9 representatives\n", "# comparing sequences from 17118 to 17159\n", "....---------- new table with 10 representatives\n", "# comparing sequences from 17159 to 17199\n", "....---------- new table with 12 representatives\n", "# comparing sequences from 17199 to 17238\n", "......---------- new table with 10 representatives\n", "# comparing sequences from 17238 to 17276\n", ".......---------- new table with 12 representatives\n", "# comparing sequences from 17276 to 17313\n", "...---------- new table with 8 representatives\n", "# comparing sequences from 17313 to 17349\n", "...---------- new table with 9 representatives\n", "# comparing sequences from 17349 to 17384\n", "...---------- new table with 7 representatives\n", "# comparing sequences from 17384 to 17418\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 17418 to 17452\n", ".....---------- new table with 8 representatives\n", "# comparing sequences from 17452 to 17485\n", "....---------- new table with 12 representatives\n", "# comparing sequences from 17485 to 17517\n", "..---------- new table with 5 representatives\n", "# comparing sequences from 17517 to 17548\n", "....---------- new table with 8 representatives\n", "# comparing sequences from 17548 to 17579\n", "..---------- new table with 8 representatives\n", "# comparing sequences from 17579 to 17609\n", ".....---------- new table with 9 representatives\n", "# comparing sequences from 17609 to 17638\n", "...---------- new table with 6 representatives\n", "# comparing sequences from 17638 to 17666\n", "....---------- new table with 8 representatives\n", "# comparing sequences from 17666 to 17694\n", ".....---------- new table with 8 representatives\n", "# comparing sequences from 17694 to 17721\n", "96.0%---------- new table with 3 representatives\n", "# comparing sequences from 17721 to 17747\n", "..---------- new table with 5 representatives\n", "# comparing sequences from 17747 to 17773\n", ".....---------- new table with 7 representatives\n", "# comparing sequences from 17773 to 17798\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17798 to 17823\n", ".---------- new table with 4 representatives\n", "# comparing sequences from 17823 to 17847\n", "...---------- new table with 4 representatives\n", "# comparing sequences from 17847 to 17870\n", "..---------- new table with 3 representatives\n", "# comparing sequences from 17870 to 18850\n", "...---------- new table with 186 representatives\n", "\n", " 18850 finished 10000 clusters\n", "\n", "Apprixmated maximum memory consumption: 534M\n", "writing new database\n", "writing clustering information\n", "program completed !\n", "\n", "Total CPU time 52.55\n", "[bwa_index] Pack FASTA... 0.01 sec\n", "[bwa_index] Construct BWT for the packed sequence...\n", "[bwa_index] 0.16 seconds elapse.\n", "[bwa_index] Update BWT... 0.00 sec\n", "[bwa_index] Pack forward-only FASTA... 0.02 sec\n", "[bwa_index] Construct SA from BWT and Occ... 0.08 sec\n", "[main] Version: 0.7.15-r1142-dirty\n", "[main] CMD: bwa index reference.fasta\n", "[main] Real time: 0.327 sec; CPU: 0.286 sec\n", "Using BWA to map reads.\n", "Creating alignment intervals\n", "Using FreeBayes to call SNPs\n", "Using VCFtools to parse SNPS.vcf for SNPs that are called in at least 90% of individuals\n", "Doing - lo\n", " New Assembly: simlo\n", "ipyrad -p params-simlo.txt -s 1\n", "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.4.0]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: simlo\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Demultiplexing fastq data to Samples\n", " [####################] 100% chunking large files | 0:00:00 \n", " [####################] 100% sorting reads | 0:00:45 \n", " [####################] 100% writing/compressing | 0:00:02 \n", "\n", "\n", "\n", "real\t0m58.457s\n", "user\t0m2.363s\n", "sys\t0m0.568s\n", "Renaming ./3J_0_R1_.fastq.gz -> Pop3_SampleJ.F.fq.gz\n", "Renaming ./3I_0_R1_.fastq.gz -> Pop3_SampleI.F.fq.gz\n", "Renaming ./1B_0_R1_.fastq.gz -> Pop1_SampleB.F.fq.gz\n", "Renaming ./2E_0_R1_.fastq.gz -> Pop2_SampleE.F.fq.gz\n", "Renaming ./2H_0_R1_.fastq.gz -> Pop2_SampleH.F.fq.gz\n", "Renaming ./1A_0_R1_.fastq.gz -> Pop1_SampleA.F.fq.gz\n", "Renaming ./2F_0_R1_.fastq.gz -> Pop2_SampleF.F.fq.gz\n", "Renaming ./3K_0_R1_.fastq.gz -> Pop3_SampleK.F.fq.gz\n", "Renaming ./1D_0_R1_.fastq.gz -> Pop1_SampleD.F.fq.gz\n", "Renaming ./2G_0_R1_.fastq.gz -> Pop2_SampleG.F.fq.gz\n", "Renaming ./3L_0_R1_.fastq.gz -> Pop3_SampleL.F.fq.gz\n", "Renaming ./1C_0_R1_.fastq.gz -> Pop1_SampleC.F.fq.gz\n", "export LD_LIBRARY_PATH=/home/iovercast/manuscript-analysis/dDocent//freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; export PATH=/home/iovercast/manuscript-analysis/dDocent/:$PATH; dDocent simlo-config.txt\n", "dDocent 2.24 \n", "\n", "Contact jpuritz@gmail.com with any problems \n", "\n", " \n", "Checking for required software\n", "\n", "All required software is installed!\n", "dDocent will require input during the assembly stage. Please wait until prompt says it is safe to move program to the background.\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "Removing the _1 character and replacing with /1 in the name of every sequence\n", "Trimming reads and simultaneously assembling reference sequences\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "sort: write failed: standard output: Broken pipe\n", "sort: write error\n", "\f", " \n", " Number of Unique Sequences with More than X Coverage (Counted within individuals) \n", " \n", " 138000 +-+---------************+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " + + ***********************************+ + + + + \n", " 136000 +-+ ********* +-+ \n", " | *** | \n", " | ***** | \n", " 134000 +-+ *** +-+ \n", " | *** | \n", " 132000 +-+ * +-+ \n", " | *** | \n", " 130000 +-+ ** +-+ \n", " | ** | \n", " 128000 +-+ ** +-+ \n", " | ** | \n", " | * | \n", " 126000 +-+ *** +-+ \n", " | * | \n", " 124000 +-+ ** +-+ \n", " | * | \n", " 122000 +-+ ** +-+ \n", " | * | \n", " | **| \n", " 120000 +-+ +-+ \n", " + + + + + + + + + * \n", " 118000 +-+---------+-----------+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " 1 2 3 4 5 6 7 8 9 10 \n", " Coverage \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. In essence, you are picking a minimum (within individual) coverage level for a read (allele) to be used in the reference assembly\n", "\f", " \n", " Number of Unique Sequences present in more than X Individuals \n", " \n", " 20000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " + + + + + + + + + \n", " ****** | \n", " | *** | \n", " 18000 +-+ *** +-+ \n", " | **** | \n", " | *** | \n", " | *** | \n", " 16000 +-+ **** +-+ \n", " | * | \n", " | ****** | \n", " 14000 +-+ **** +-+ \n", " | **** | \n", " | **** | \n", " | **** | \n", " 12000 +-+ **** +-+ \n", " | * | \n", " | *********** | \n", " | ****** | \n", " 10000 +-+ ******* +-+ \n", " | *** | \n", " | ***************************| \n", " + + + + + + + + * \n", " 8000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " 2 2.5 3 3.5 4 4.5 5 5.5 6 \n", " Number of Individuals \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. Pick point right before the assymptote. A good starting cutoff might be 10% of the total number of individuals\n", "At this point, all configuration information has been entered and dDocent may take several hours to run.\n", "It is recommended that you move this script to a background operation and disable terminal input and output.\n", "All data and logfiles will still be recorded.\n", "To do this:\n", "Press control and Z simultaneously\n", "Type 'bg' without the quotes and press enter\n", "Type 'disown -h' again without the quotes and press enter\n", "\n", "Now sit back, relax, and wait for your analysis to finish.\n", "TrimmomaticSE: Started with arguments: -threads 40 -phred33 uniq.fq uniq.fq1 ILLUMINACLIP:/home/iovercast/manuscript-analysis/dDocent/TruSeq2-PE.fa:2:30:10 MINLEN:68\n", "Using PrefixPair: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT' and 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTAATGATACGGCGACCACCGAGATCTACAC'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTCAAGCAGAAGACGGCATACGA'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGCCGTATCATT'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGGTTCAGCAGGAATGCCGAGACCGATCTCGTATGCCGTCTTCTGCTTG'\n", "Using Long Clipping Sequence: 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT'\n", "ILLUMINACLIP: Using 1 prefix pairs, 6 forward/reverse sequences, 0 forward only sequences, 0 reverse only sequences\n", "Input Reads: 18847 Surviving: 18847 (100.00%) Dropped: 0 (0.00%)\n", "TrimmomaticSE: Completed successfully\n", "================================================================\n", "Program: CD-HIT, V4.6 (+OpenMP), Sep 21 2016, 20:52:31\n", "Command: cd-hit-est -i totalover.fasta -o\n", " reference.fasta.original -M 0 -T 0 -c 0.85\n", "\n", "Started: Sat Oct 1 18:04:24 2016\n", "================================================================\n", " Output \n", "----------------------------------------------------------------\n", "total number of CPUs in the system is 40\n", "Actual number of CPUs to be used: 40\n", "\n", "total seq: 18847\n", "longest and shortest : 91 and 91\n", "Total letters: 1715077\n", "Sequences have been sorted\n", "\n", "Approximated minimal memory consumption:\n", "Sequence : 4M\n", "Buffer : 40 X 12M = 491M\n", "Table : 2 X 17M = 34M\n", "Miscellaneous : 4M\n", "Total : 534M\n", "\n", "Table limit with the given memory limit:\n", "Max number of representatives: 366227\n", "Max number of word counting entries: 8747853\n", "\n", "# comparing sequences from 0 to 448\n", "---------- new table with 442 representatives\n", "# comparing sequences from 448 to 886\n", "---------- 334 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 552 to 987\n", "---------- 331 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 656 to 1089\n", "---------- 327 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 762 to 1192\n", "---------- 194 remaining sequences to the next cycle\n", "---------- new table with 222 representatives\n", "# comparing sequences from 998 to 1422\n", "---------- 318 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1104 to 1526\n", "---------- 315 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1211 to 1630\n", "---------- 316 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1314 to 1731\n", "---------- 290 remaining sequences to the next cycle\n", "---------- new table with 120 representatives\n", "# comparing sequences from 1441 to 1855\n", "---------- 301 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1554 to 1965\n", "---------- 183 remaining sequences to the next cycle\n", "---------- new table with 202 representatives\n", "# comparing sequences from 1782 to 2188\n", "---------- 291 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 1897 to 2300\n", "---------- 195 remaining sequences to the next cycle\n", "---------- new table with 184 representatives\n", "# comparing sequences from 2105 to 2503\n", "---------- 151 remaining sequences to the next cycle\n", "---------- new table with 212 representatives\n", "# comparing sequences from 2352 to 2744\n", "---------- 108 remaining sequences to the next cycle\n", "---------- new table with 246 representatives\n", "# comparing sequences from 2636 to 3021\n", "---------- 114 remaining sequences to the next cycle\n", "---------- new table with 219 representatives\n", "# comparing sequences from 2907 to 3286\n", "---------- 87 remaining sequences to the next cycle\n", "---------- new table with 241 representatives\n", "# comparing sequences from 3199 to 3571\n", "---------- 243 remaining sequences to the next cycle\n", "---------- new table with 105 representatives\n", "# comparing sequences from 3328 to 3697\n", "---------- 145 remaining sequences to the next cycle\n", "---------- new table with 181 representatives\n", "# comparing sequences from 3552 to 3916\n", "---------- 248 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 3668 to 4029\n", "---------- 235 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 3794 to 4152\n", "---------- 152 remaining sequences to the next cycle\n", "---------- new table with 153 representatives\n", "# comparing sequences from 4000 to 4353\n", "---------- 131 remaining sequences to the next cycle\n", "---------- new table with 164 representatives\n", "# comparing sequences from 4222 to 4570\n", "---------- 86 remaining sequences to the next cycle\n", "---------- new table with 187 representatives\n", "# comparing sequences from 4484 to 4825\n", "---------- 112 remaining sequences to the next cycle\n", "---------- new table with 164 representatives\n", "# comparing sequences from 4713 to 5049\n", "---------- 134 remaining sequences to the next cycle\n", "---------- new table with 149 representatives\n", "# comparing sequences from 4915 to 5246\n", "---------- 191 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5055 to 5383\n", "---------- 184 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5199 to 5523\n", "---------- 184 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5339 to 5660\n", "---------- 141 remaining sequences to the next cycle\n", "---------- new table with 129 representatives\n", "# comparing sequences from 5519 to 5836\n", "---------- 175 remaining sequences to the next cycle\n", "---------- new table with 103 representatives\n", "# comparing sequences from 5661 to 5974\n", "---------- 166 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5808 to 6118\n", "---------- 147 remaining sequences to the next cycle\n", "---------- new table with 103 representatives\n", "# comparing sequences from 5971 to 6277\n", "---------- 143 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6134 to 6436\n", "---------- 143 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6293 to 6591\n", "---------- 135 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6456 to 6751\n", "---------- 134 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6617 to 6908\n", "---------- 128 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6780 to 7067\n", "---------- 126 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6941 to 7224\n", "---------- 115 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7109 to 7388\n", "---------- 120 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7268 to 7543\n", "---------- 106 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7437 to 7708\n", "---------- 100 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7608 to 7875\n", "---------- 99 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7776 to 8039\n", "---------- 76 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7963 to 8222\n", "---------- 87 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8135 to 8390\n", "---------- 50 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8340 to 8590\n", "---------- 59 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8531 to 8776\n", "---------- 58 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8718 to 8959\n", "---------- 32 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8927 to 9163\n", "---------- 28 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9135 to 9366\n", "---------- 45 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9321 to 9547\n", "---------- 36 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9511 to 9733\n", "---------- 28 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9705 to 9922\n", "---------- 6 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9916 to 10128\n", "---------- 4 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 10124 to 10331\n", ".......---------- new table with 99 representatives\n", "# comparing sequences from 10331 to 10533\n", "---------- 6 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 10527 to 10725\n", "...........---------- new table with 97 representatives\n", "# comparing sequences from 10725 to 10918\n", ".........---------- new table with 86 representatives\n", "# comparing sequences from 10918 to 11106\n", "......---------- new table with 70 representatives\n", "# comparing sequences from 11106 to 11290\n", "............---------- new table with 82 representatives\n", "# comparing sequences from 11290 to 11469\n", "........---------- new table with 74 representatives\n", "# comparing sequences from 11469 to 11644\n", "......---------- new table with 76 representatives\n", "# comparing sequences from 11644 to 11815\n", ".........---------- new table with 75 representatives\n", "# comparing sequences from 11815 to 11982\n", "........---------- new table with 77 representatives\n", "# comparing sequences from 11982 to 12145\n", ".....---------- new table with 56 representatives\n", "# comparing sequences from 12145 to 12304\n", "......---------- new table with 58 representatives\n", "# comparing sequences from 12304 to 12459\n", ".............---------- new table with 69 representatives\n", "# comparing sequences from 12459 to 12611\n", "....---------- new table with 55 representatives\n", "# comparing sequences from 12611 to 12759\n", "......---------- new table with 49 representatives\n", "# comparing sequences from 12759 to 12903\n", ".....---------- new table with 49 representatives\n", "# comparing sequences from 12903 to 13044\n", "..........---------- new table with 54 representatives\n", "# comparing sequences from 13044 to 13182\n", "..........---------- new table with 52 representatives\n", "# comparing sequences from 13182 to 13316\n", ".....---------- new table with 39 representatives\n", "# comparing sequences from 13316 to 13447\n", "....---------- new table with 38 representatives\n", "# comparing sequences from 13447 to 13575\n", ".....---------- new table with 47 representatives\n", "# comparing sequences from 13575 to 13700\n", ".....---------- new table with 41 representatives\n", "# comparing sequences from 13700 to 13822\n", ".......---------- new table with 35 representatives\n", "# comparing sequences from 13822 to 13941\n", ".....---------- new table with 30 representatives\n", "# comparing sequences from 13941 to 14057\n", "............---------- new table with 42 representatives\n", "# comparing sequences from 14057 to 14171\n", ".........---------- new table with 35 representatives\n", "# comparing sequences from 14171 to 14282\n", "......---------- new table with 36 representatives\n", "# comparing sequences from 14282 to 14390\n", ".........---------- new table with 38 representatives\n", "# comparing sequences from 14390 to 14496\n", ".....---------- new table with 36 representatives\n", "# comparing sequences from 14496 to 14599\n", ".......---------- new table with 34 representatives\n", "# comparing sequences from 14599 to 14700\n", "........---------- new table with 27 representatives\n", "# comparing sequences from 14700 to 14798\n", "....---------- new table with 33 representatives\n", "# comparing sequences from 14798 to 14894\n", ".......---------- new table with 31 representatives\n", "# comparing sequences from 14894 to 14988\n", ".......---------- new table with 28 representatives\n", "# comparing sequences from 14988 to 15079\n", ".......---------- new table with 28 representatives\n", "# comparing sequences from 15079 to 15168\n", "........---------- new table with 22 representatives\n", "# comparing sequences from 15168 to 15255\n", ".......---------- new table with 25 representatives\n", "# comparing sequences from 15255 to 15340\n", "..........---------- new table with 30 representatives\n", "# comparing sequences from 15340 to 15423\n", ".......---------- new table with 27 representatives\n", "# comparing sequences from 15423 to 15504\n", "......---------- new table with 22 representatives\n", "# comparing sequences from 15504 to 15583\n", "........---------- new table with 23 representatives\n", "# comparing sequences from 15583 to 15660\n", ".....---------- new table with 21 representatives\n", "# comparing sequences from 15660 to 15735\n", "........---------- new table with 20 representatives\n", "# comparing sequences from 15735 to 15809\n", ".......---------- new table with 22 representatives\n", "# comparing sequences from 15809 to 15881\n", "....---------- new table with 18 representatives\n", "# comparing sequences from 15881 to 15951\n", "......---------- new table with 19 representatives\n", "# comparing sequences from 15951 to 16019\n", "..---------- new table with 13 representatives\n", "# comparing sequences from 16019 to 16086\n", ".....---------- new table with 15 representatives\n", "# comparing sequences from 16086 to 16151\n", ".......---------- new table with 17 representatives\n", "# comparing sequences from 16151 to 16215\n", "....---------- new table with 19 representatives\n", "# comparing sequences from 16215 to 16277\n", "......---------- new table with 15 representatives\n", "# comparing sequences from 16277 to 16338\n", "....---------- new table with 15 representatives\n", "# comparing sequences from 16338 to 16397\n", "...---------- new table with 17 representatives\n", "# comparing sequences from 16397 to 16455\n", "..---------- new table with 17 representatives\n", "# comparing sequences from 16455 to 16511\n", ".....---------- new table with 11 representatives\n", "# comparing sequences from 16511 to 16566\n", "99.9%---------- new table with 6 representatives\n", "# comparing sequences from 16566 to 16620\n", ".........---------- new table with 14 representatives\n", "# comparing sequences from 16620 to 16673\n", ".....---------- new table with 12 representatives\n", "# comparing sequences from 16673 to 16724\n", ".....---------- new table with 16 representatives\n", "# comparing sequences from 16724 to 16774\n", "...---------- new table with 13 representatives\n", "# comparing sequences from 16774 to 16823\n", "....---------- new table with 11 representatives\n", "# comparing sequences from 16823 to 16871\n", "....---------- new table with 11 representatives\n", "# comparing sequences from 16871 to 16918\n", "......---------- new table with 11 representatives\n", "# comparing sequences from 16918 to 16963\n", "..---------- new table with 8 representatives\n", "# comparing sequences from 16963 to 17007\n", "...---------- new table with 10 representatives\n", "# comparing sequences from 17007 to 17050\n", "....---------- new table with 9 representatives\n", "# comparing sequences from 17050 to 17092\n", "...---------- new table with 14 representatives\n", "# comparing sequences from 17092 to 17133\n", "...---------- new table with 6 representatives\n", "# comparing sequences from 17133 to 17173\n", "......---------- new table with 13 representatives\n", "# comparing sequences from 17173 to 17212\n", ".....---------- new table with 12 representatives\n", "# comparing sequences from 17212 to 17250\n", ".....---------- new table with 9 representatives\n", "# comparing sequences from 17250 to 17288\n", ".......---------- new table with 10 representatives\n", "# comparing sequences from 17288 to 17325\n", ".......---------- new table with 11 representatives\n", "# comparing sequences from 17325 to 17361\n", "....---------- new table with 8 representatives\n", "# comparing sequences from 17361 to 17396\n", "......---------- new table with 11 representatives\n", "# comparing sequences from 17396 to 17430\n", "...---------- new table with 9 representatives\n", "# comparing sequences from 17430 to 17463\n", "...---------- new table with 11 representatives\n", "# comparing sequences from 17463 to 17495\n", "....---------- new table with 7 representatives\n", "# comparing sequences from 17495 to 17527\n", "....---------- new table with 6 representatives\n", "# comparing sequences from 17527 to 17558\n", "...---------- new table with 7 representatives\n", "# comparing sequences from 17558 to 17588\n", "......---------- new table with 11 representatives\n", "# comparing sequences from 17588 to 17617\n", "..---------- new table with 5 representatives\n", "# comparing sequences from 17617 to 17646\n", ".....---------- new table with 8 representatives\n", "# comparing sequences from 17646 to 17674\n", "......---------- new table with 10 representatives\n", "# comparing sequences from 17674 to 17701\n", ".---------- new table with 2 representatives\n", "# comparing sequences from 17701 to 17728\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17728 to 17754\n", ".....---------- new table with 7 representatives\n", "# comparing sequences from 17754 to 17780\n", "....---------- new table with 7 representatives\n", "# comparing sequences from 17780 to 17805\n", "...---------- new table with 3 representatives\n", "# comparing sequences from 17805 to 17829\n", ".---------- new table with 5 representatives\n", "# comparing sequences from 17829 to 17853\n", ".---------- new table with 2 representatives\n", "# comparing sequences from 17853 to 18847\n", "....---------- new table with 190 representatives\n", "\n", " 18847 finished 10000 clusters\n", "\n", "Apprixmated maximum memory consumption: 534M\n", "writing new database\n", "writing clustering information\n", "program completed !\n", "\n", "Total CPU time 55.78\n", "[bwa_index] Pack FASTA... 0.01 sec\n", "[bwa_index] Construct BWT for the packed sequence...\n", "[bwa_index] 0.16 seconds elapse.\n", "[bwa_index] Update BWT... 0.01 sec\n", "[bwa_index] Pack forward-only FASTA... 0.01 sec\n", "[bwa_index] Construct SA from BWT and Occ... 0.09 sec\n", "[main] Version: 0.7.15-r1142-dirty\n", "[main] CMD: bwa index reference.fasta\n", "[main] Real time: 0.338 sec; CPU: 0.299 sec\n", "Using BWA to map reads.\n", "Creating alignment intervals\n", "Using FreeBayes to call SNPs\n", "Using VCFtools to parse SNPS.vcf for SNPs that are called in at least 90% of individuals\n", "Doing - hi\n", " New Assembly: simhi\n", "ipyrad -p params-simhi.txt -s 1\n", "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.4.0]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: simhi\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Demultiplexing fastq data to Samples\n", " [####################] 100% chunking large files | 0:00:00 \n", " [####################] 100% sorting reads | 0:00:45 \n", " [####################] 100% writing/compressing | 0:00:02 \n", "\n", "\n", "\n", "real\t0m57.199s\n", "user\t0m1.775s\n", "sys\t0m0.522s\n", "Renaming ./3K_0_R1_.fastq.gz -> Pop3_SampleK.F.fq.gz\n", "Renaming ./2G_0_R1_.fastq.gz -> Pop2_SampleG.F.fq.gz\n", "Renaming ./2E_0_R1_.fastq.gz -> Pop2_SampleE.F.fq.gz\n", "Renaming ./3L_0_R1_.fastq.gz -> Pop3_SampleL.F.fq.gz\n", "Renaming ./1A_0_R1_.fastq.gz -> Pop1_SampleA.F.fq.gz\n", "Renaming ./2H_0_R1_.fastq.gz -> Pop2_SampleH.F.fq.gz\n", "Renaming ./3J_0_R1_.fastq.gz -> Pop3_SampleJ.F.fq.gz\n", "Renaming ./1B_0_R1_.fastq.gz -> Pop1_SampleB.F.fq.gz\n", "Renaming ./1D_0_R1_.fastq.gz -> Pop1_SampleD.F.fq.gz\n", "Renaming ./2F_0_R1_.fastq.gz -> Pop2_SampleF.F.fq.gz\n", "Renaming ./3I_0_R1_.fastq.gz -> Pop3_SampleI.F.fq.gz\n", "Renaming ./1C_0_R1_.fastq.gz -> Pop1_SampleC.F.fq.gz\n", "export LD_LIBRARY_PATH=/home/iovercast/manuscript-analysis/dDocent//freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; export PATH=/home/iovercast/manuscript-analysis/dDocent/:$PATH; dDocent simhi-config.txt\n", "dDocent 2.24 \n", "\n", "Contact jpuritz@gmail.com with any problems \n", "\n", " \n", "Checking for required software\n", "\n", "All required software is installed!\n", "dDocent will require input during the assembly stage. Please wait until prompt says it is safe to move program to the background.\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "Removing the _1 character and replacing with /1 in the name of every sequence\n", "Trimming reads and simultaneously assembling reference sequences\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "sort: write failed: standard output: Broken pipe\n", "sort: write error\n", "\f", " \n", " Number of Unique Sequences with More than X Coverage (Counted within individuals) \n", " \n", " 138000 +-+---------************+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " + + ***********************************+ + + + + \n", " 136000 +-+ ********* +-+ \n", " | *** | \n", " | ***** | \n", " 134000 +-+ *** +-+ \n", " | *** | \n", " 132000 +-+ * +-+ \n", " | *** | \n", " 130000 +-+ ** +-+ \n", " | ** | \n", " 128000 +-+ ** +-+ \n", " | ** | \n", " | * | \n", " 126000 +-+ *** +-+ \n", " | * | \n", " 124000 +-+ ** +-+ \n", " | * | \n", " 122000 +-+ ** +-+ \n", " | * | \n", " | **| \n", " 120000 +-+ +-+ \n", " + + + + + + + + + * \n", " 118000 +-+---------+-----------+-----------+-----------+----------+-----------+-----------+-----------+---------+-+ \n", " 1 2 3 4 5 6 7 8 9 10 \n", " Coverage \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. In essence, you are picking a minimum (within individual) coverage level for a read (allele) to be used in the reference assembly\n", "\f", " \n", " Number of Unique Sequences present in more than X Individuals \n", " \n", " 20000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " + + + + + + + + + \n", " ****** | \n", " | *** | \n", " 18000 +-+ *** +-+ \n", " | **** | \n", " | *** | \n", " | *** | \n", " 16000 +-+ **** +-+ \n", " | * | \n", " | ******* | \n", " 14000 +-+ ***** +-+ \n", " | **** | \n", " | ***** | \n", " | **** | \n", " 12000 +-+ ** +-+ \n", " | ********* | \n", " | ***** | \n", " | ***** | \n", " 10000 +-+ ****** +-+ \n", " | ** | \n", " | ***************************| \n", " + + + + + + + + * \n", " 8000 +-+-----------+------------+-------------+------------+-------------+------------+-------------+----------+-+ \n", " 2 2.5 3 3.5 4 4.5 5 5.5 6 \n", " Number of Individuals \n", " \n", "\u0007\u0007\u0007Please choose data cutoff. Pick point right before the assymptote. A good starting cutoff might be 10% of the total number of individuals\n", "At this point, all configuration information has been entered and dDocent may take several hours to run.\n", "It is recommended that you move this script to a background operation and disable terminal input and output.\n", "All data and logfiles will still be recorded.\n", "To do this:\n", "Press control and Z simultaneously\n", "Type 'bg' without the quotes and press enter\n", "Type 'disown -h' again without the quotes and press enter\n", "\n", "Now sit back, relax, and wait for your analysis to finish.\n", "TrimmomaticSE: Started with arguments: -threads 40 -phred33 uniq.fq uniq.fq1 ILLUMINACLIP:/home/iovercast/manuscript-analysis/dDocent/TruSeq2-PE.fa:2:30:10 MINLEN:68\n", "Using PrefixPair: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT' and 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTAATGATACGGCGACCACCGAGATCTACAC'\n", "Using Long Clipping Sequence: 'TTTTTTTTTTCAAGCAGAAGACGGCATACGA'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTAGATCTCGGTGGTCGCCGTATCATT'\n", "Using Long Clipping Sequence: 'AGATCGGAAGAGCGGTTCAGCAGGAATGCCGAGACCGATCTCGTATGCCGTCTTCTGCTTG'\n", "Using Long Clipping Sequence: 'CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT'\n", "Using Long Clipping Sequence: 'AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT'\n", "ILLUMINACLIP: Using 1 prefix pairs, 6 forward/reverse sequences, 0 forward only sequences, 0 reverse only sequences\n", "Input Reads: 18867 Surviving: 18867 (100.00%) Dropped: 0 (0.00%)\n", "TrimmomaticSE: Completed successfully\n", "================================================================\n", "Program: CD-HIT, V4.6 (+OpenMP), Sep 21 2016, 20:52:31\n", "Command: cd-hit-est -i totalover.fasta -o\n", " reference.fasta.original -M 0 -T 0 -c 0.85\n", "\n", "Started: Sat Oct 1 18:07:19 2016\n", "================================================================\n", " Output \n", "----------------------------------------------------------------\n", "total number of CPUs in the system is 40\n", "Actual number of CPUs to be used: 40\n", "\n", "total seq: 18867\n", "longest and shortest : 91 and 91\n", "Total letters: 1716897\n", "Sequences have been sorted\n", "\n", "Approximated minimal memory consumption:\n", "Sequence : 4M\n", "Buffer : 40 X 12M = 491M\n", "Table : 2 X 17M = 34M\n", "Miscellaneous : 4M\n", "Total : 534M\n", "\n", "Table limit with the given memory limit:\n", "Max number of representatives: 366227\n", "Max number of word counting entries: 8747853\n", "\n", "# comparing sequences from 0 to 449\n", "---------- new table with 444 representatives\n", "# comparing sequences from 449 to 887\n", "---------- 334 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 553 to 989\n", "---------- 241 remaining sequences to the next cycle\n", "---------- new table with 185 representatives\n", "# comparing sequences from 748 to 1179\n", "---------- 239 remaining sequences to the next cycle\n", "---------- new table with 181 representatives\n", "# comparing sequences from 940 to 1366\n", "---------- 206 remaining sequences to the next cycle\n", "---------- new table with 205 representatives\n", "# comparing sequences from 1160 to 1581\n", "---------- 205 remaining sequences to the next cycle\n", "---------- new table with 202 representatives\n", "# comparing sequences from 1376 to 1792\n", "---------- 200 remaining sequences to the next cycle\n", "---------- new table with 197 representatives\n", "# comparing sequences from 1592 to 2003\n", "---------- 195 remaining sequences to the next cycle\n", "---------- new table with 196 representatives\n", "# comparing sequences from 1808 to 2214\n", "---------- 172 remaining sequences to the next cycle\n", "---------- new table with 204 representatives\n", "# comparing sequences from 2042 to 2442\n", "---------- 215 remaining sequences to the next cycle\n", "---------- new table with 160 representatives\n", "# comparing sequences from 2227 to 2623\n", "---------- 188 remaining sequences to the next cycle\n", "---------- new table with 177 representatives\n", "# comparing sequences from 2435 to 2826\n", "---------- 204 remaining sequences to the next cycle\n", "---------- new table with 162 representatives\n", "# comparing sequences from 2622 to 3008\n", "---------- 177 remaining sequences to the next cycle\n", "---------- new table with 171 representatives\n", "# comparing sequences from 2831 to 3212\n", "---------- 91 remaining sequences to the next cycle\n", "---------- new table with 235 representatives\n", "# comparing sequences from 3121 to 3495\n", "---------- 122 remaining sequences to the next cycle\n", "---------- new table with 207 representatives\n", "# comparing sequences from 3373 to 3741\n", "---------- 248 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 3493 to 3859\n", "---------- 113 remaining sequences to the next cycle\n", "---------- new table with 211 representatives\n", "# comparing sequences from 3746 to 4106\n", "---------- 115 remaining sequences to the next cycle\n", "---------- new table with 187 representatives\n", "# comparing sequences from 3991 to 4345\n", "80.8%---------- new table with 258 representatives\n", "# comparing sequences from 4345 to 4690\n", "---------- 127 remaining sequences to the next cycle\n", "---------- new table with 158 representatives\n", "# comparing sequences from 4563 to 4903\n", "---------- 154 remaining sequences to the next cycle\n", "---------- new table with 142 representatives\n", "# comparing sequences from 4749 to 5085\n", "---------- 199 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 4886 to 5218\n", "---------- 189 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5029 to 5358\n", "---------- 183 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 5175 to 5501\n", "100.0%---------- new table with 230 representatives\n", "# comparing sequences from 5501 to 5819\n", "86.1%---------- new table with 219 representatives\n", "# comparing sequences from 5819 to 6129\n", "---------- 39 remaining sequences to the next cycle\n", "---------- new table with 166 representatives\n", "# comparing sequences from 6090 to 6394\n", "---------- 52 remaining sequences to the next cycle\n", "---------- new table with 155 representatives\n", "# comparing sequences from 6342 to 6640\n", "---------- 140 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 6500 to 6794\n", "---------- 77 remaining sequences to the next cycle\n", "---------- new table with 131 representatives\n", "# comparing sequences from 6717 to 7006\n", "---------- 41 remaining sequences to the next cycle\n", "---------- new table with 145 representatives\n", "# comparing sequences from 6965 to 7248\n", "---------- 64 remaining sequences to the next cycle\n", "---------- new table with 142 representatives\n", "# comparing sequences from 7184 to 7462\n", "---------- 52 remaining sequences to the next cycle\n", "---------- new table with 132 representatives\n", "# comparing sequences from 7410 to 7682\n", "---------- 96 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7586 to 7854\n", "---------- 97 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7757 to 8021\n", "---------- 93 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 7928 to 8188\n", "---------- 90 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8098 to 8354\n", "---------- 52 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8302 to 8553\n", "---------- 77 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8476 to 8723\n", "---------- 44 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8679 to 8921\n", "---------- 33 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 8888 to 9125\n", "---------- 50 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9075 to 9308\n", "---------- 51 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9257 to 9485\n", "99.9%---------- new table with 115 representatives\n", "# comparing sequences from 9485 to 9708\n", "---------- 21 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9687 to 9905\n", "---------- 11 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 9894 to 10107\n", "........---------- new table with 97 representatives\n", "# comparing sequences from 10107 to 10315\n", "...........---------- new table with 94 representatives\n", "# comparing sequences from 10315 to 10518\n", "---------- 9 remaining sequences to the next cycle\n", "---------- new table with 100 representatives\n", "# comparing sequences from 10509 to 10708\n", ".............---------- new table with 94 representatives\n", "# comparing sequences from 10708 to 10902\n", ".............---------- new table with 92 representatives\n", "# comparing sequences from 10902 to 11091\n", ".......---------- new table with 73 representatives\n", "# comparing sequences from 11091 to 11276\n", "..........---------- new table with 84 representatives\n", "# comparing sequences from 11276 to 11456\n", "..........---------- new table with 79 representatives\n", "# comparing sequences from 11456 to 11632\n", "....---------- new table with 62 representatives\n", "# comparing sequences from 11632 to 11804\n", ".............---------- new table with 85 representatives\n", "# comparing sequences from 11804 to 11972\n", ".........---------- new table with 69 representatives\n", "# comparing sequences from 11972 to 12136\n", "....---------- new table with 61 representatives\n", "# comparing sequences from 12136 to 12296\n", ".....---------- new table with 55 representatives\n", "# comparing sequences from 12296 to 12452\n", "...........---------- new table with 69 representatives\n", "# comparing sequences from 12452 to 12604\n", ".......---------- new table with 61 representatives\n", "# comparing sequences from 12604 to 12753\n", ".......---------- new table with 50 representatives\n", "# comparing sequences from 12753 to 12898\n", "...---------- new table with 50 representatives\n", "# comparing sequences from 12898 to 13040\n", "......---------- new table with 58 representatives\n", "# comparing sequences from 13040 to 13178\n", "........---------- new table with 53 representatives\n", "# comparing sequences from 13178 to 13313\n", "......---------- new table with 39 representatives\n", "# comparing sequences from 13313 to 13445\n", "........---------- new table with 42 representatives\n", "# comparing sequences from 13445 to 13574\n", ".....---------- new table with 40 representatives\n", "# comparing sequences from 13574 to 13700\n", ".......---------- new table with 43 representatives\n", "# comparing sequences from 13700 to 13823\n", ".....---------- new table with 37 representatives\n", "# comparing sequences from 13823 to 13943\n", "....---------- new table with 33 representatives\n", "# comparing sequences from 13943 to 14060\n", ".......---------- new table with 37 representatives\n", "# comparing sequences from 14060 to 14174\n", "......---------- new table with 36 representatives\n", "# comparing sequences from 14174 to 14285\n", ".....---------- new table with 42 representatives\n", "# comparing sequences from 14285 to 14394\n", ".....---------- new table with 37 representatives\n", "# comparing sequences from 14394 to 14500\n", "....---------- new table with 34 representatives\n", "# comparing sequences from 14500 to 14603\n", "....---------- new table with 30 representatives\n", "# comparing sequences from 14603 to 14704\n", "........---------- new table with 33 representatives\n", "# comparing sequences from 14704 to 14803\n", "..........---------- new table with 34 representatives\n", "# comparing sequences from 14803 to 14899\n", "......---------- new table with 30 representatives\n", "# comparing sequences from 14899 to 14993\n", ".......---------- new table with 30 representatives\n", "# comparing sequences from 14993 to 15085\n", "....---------- new table with 26 representatives\n", "# comparing sequences from 15085 to 15175\n", ".....---------- new table with 25 representatives\n", "# comparing sequences from 15175 to 15262\n", ".....---------- new table with 24 representatives\n", "# comparing sequences from 15262 to 15347\n", "...---------- new table with 27 representatives\n", "# comparing sequences from 15347 to 15430\n", ".....---------- new table with 25 representatives\n", "# comparing sequences from 15430 to 15511\n", "...---------- new table with 22 representatives\n", "# comparing sequences from 15511 to 15590\n", ".......---------- new table with 22 representatives\n", "# comparing sequences from 15590 to 15668\n", "......---------- new table with 22 representatives\n", "# comparing sequences from 15668 to 15744\n", "......---------- new table with 20 representatives\n", "# comparing sequences from 15744 to 15818\n", "........---------- new table with 25 representatives\n", "# comparing sequences from 15818 to 15890\n", "....---------- new table with 12 representatives\n", "# comparing sequences from 15890 to 15960\n", "....---------- new table with 17 representatives\n", "# comparing sequences from 15960 to 16029\n", ".....---------- new table with 16 representatives\n", "# comparing sequences from 16029 to 16096\n", ".---------- new table with 16 representatives\n", "# comparing sequences from 16096 to 16161\n", "...---------- new table with 17 representatives\n", "# comparing sequences from 16161 to 16225\n", "......---------- new table with 18 representatives\n", "# comparing sequences from 16225 to 16287\n", "....---------- new table with 17 representatives\n", "# comparing sequences from 16287 to 16348\n", ".---------- new table with 12 representatives\n", "# comparing sequences from 16348 to 16407\n", "....---------- new table with 16 representatives\n", "# comparing sequences from 16407 to 16465\n", "...---------- new table with 14 representatives\n", "# comparing sequences from 16465 to 16522\n", ".......---------- new table with 14 representatives\n", "# comparing sequences from 16522 to 16577\n", "...---------- new table with 10 representatives\n", "# comparing sequences from 16577 to 16631\n", ".---------- new table with 14 representatives\n", "# comparing sequences from 16631 to 16684\n", "...---------- new table with 10 representatives\n", "# comparing sequences from 16684 to 16735\n", ".....---------- new table with 15 representatives\n", "# comparing sequences from 16735 to 16785\n", ".....---------- new table with 14 representatives\n", "# comparing sequences from 16785 to 16834\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 16834 to 16882\n", "....---------- new table with 13 representatives\n", "# comparing sequences from 16882 to 16929\n", ".---------- new table with 6 representatives\n", "# comparing sequences from 16929 to 16975\n", ".......---------- new table with 13 representatives\n", "# comparing sequences from 16975 to 17020\n", "....---------- new table with 9 representatives\n", "# comparing sequences from 17020 to 17063\n", "..---------- new table with 10 representatives\n", "# comparing sequences from 17063 to 17105\n", "......---------- new table with 11 representatives\n", "# comparing sequences from 17105 to 17146\n", "..---------- new table with 7 representatives\n", "# comparing sequences from 17146 to 17186\n", "...---------- new table with 12 representatives\n", "# comparing sequences from 17186 to 17226\n", ".---------- new table with 9 representatives\n", "# comparing sequences from 17226 to 17265\n", ".....---------- new table with 10 representatives\n", "# comparing sequences from 17265 to 17303\n", ".........---------- new table with 12 representatives\n", "# comparing sequences from 17303 to 17340\n", "....---------- new table with 9 representatives\n", "# comparing sequences from 17340 to 17376\n", ".....---------- new table with 8 representatives\n", "# comparing sequences from 17376 to 17411\n", ".....---------- new table with 9 representatives\n", "# comparing sequences from 17411 to 17445\n", "......---------- new table with 12 representatives\n", "# comparing sequences from 17445 to 17478\n", "...---------- new table with 11 representatives\n", "# comparing sequences from 17478 to 17511\n", ".........---------- new table with 12 representatives\n", "# comparing sequences from 17511 to 17543\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17543 to 17574\n", "...---------- new table with 8 representatives\n", "# comparing sequences from 17574 to 17604\n", "...---------- new table with 9 representatives\n", "# comparing sequences from 17604 to 17634\n", "...---------- new table with 6 representatives\n", "# comparing sequences from 17634 to 17663\n", ".---------- new table with 4 representatives\n", "# comparing sequences from 17663 to 17691\n", "....---------- new table with 11 representatives\n", "# comparing sequences from 17691 to 17719\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17719 to 17746\n", "...---------- new table with 5 representatives\n", "# comparing sequences from 17746 to 17772\n", "....---------- new table with 5 representatives\n", "# comparing sequences from 17772 to 17798\n", "...---------- new table with 7 representatives\n", "# comparing sequences from 17798 to 17823\n", "---------- new table with 3 representatives\n", "# comparing sequences from 17823 to 17847\n", "....---------- new table with 6 representatives\n", "# comparing sequences from 17847 to 17871\n", "95.8%---------- new table with 1 representatives\n", "# comparing sequences from 17871 to 18867\n", ".....---------- new table with 189 representatives\n", "\n", " 18867 finished 10000 clusters\n", "\n", "Apprixmated maximum memory consumption: 534M\n", "writing new database\n", "writing clustering information\n", "program completed !\n", "\n", "Total CPU time 53.40\n", "[bwa_index] Pack FASTA... 0.01 sec\n", "[bwa_index] Construct BWT for the packed sequence...\n", "[bwa_index] 0.24 seconds elapse.\n", "[bwa_index] Update BWT... 0.01 sec\n", "[bwa_index] Pack forward-only FASTA... 0.02 sec\n", "[bwa_index] Construct SA from BWT and Occ... 0.08 sec\n", "[main] Version: 0.7.15-r1142-dirty\n", "[main] CMD: bwa index reference.fasta\n", "[main] Real time: 0.409 sec; CPU: 0.369 sec\n", "Using BWA to map reads.\n", "Creating alignment intervals\n", "Using FreeBayes to call SNPs\n", "Using VCFtools to parse SNPS.vcf for SNPs that are called in at least 90% of individuals\n", "Doing - large\n", " New Assembly: simlarge\n", "ipyrad -p params-simlarge.txt -s 1\n", "\n", " -------------------------------------------------------------\n", " ipyrad [v.0.4.0]\n", " Interactive assembly and analysis of RAD-seq data\n", " -------------------------------------------------------------\n", " New Assembly: simlarge\n", " local compute node: [40 cores] on node001\n", "\n", " Step 1: Demultiplexing fastq data to Samples\n", " [####################] 100% chunking large files | 0:01:06 \n", " [####################] 100% sorting reads | 0:00:59 \n", " [####################] 100% writing/compressing | 0:00:11 \n", "\n", "\n", "\n", "real\t2m28.462s\n", "user\t0m2.154s\n", "sys\t0m1.422s\n", "Renaming ./3K_0_R1_.fastq.gz -> Pop3_SampleK.F.fq.gz\n", "Renaming ./2H_0_R1_.fastq.gz -> Pop2_SampleH.F.fq.gz\n", "Renaming ./1B_0_R1_.fastq.gz -> Pop1_SampleB.F.fq.gz\n", "Renaming ./3L_0_R1_.fastq.gz -> Pop3_SampleL.F.fq.gz\n", "Renaming ./1C_0_R1_.fastq.gz -> Pop1_SampleC.F.fq.gz\n", "Renaming ./1A_0_R1_.fastq.gz -> Pop1_SampleA.F.fq.gz\n", "Renaming ./2G_0_R1_.fastq.gz -> Pop2_SampleG.F.fq.gz\n", "Renaming ./2E_0_R1_.fastq.gz -> Pop2_SampleE.F.fq.gz\n", "Renaming ./3J_0_R1_.fastq.gz -> Pop3_SampleJ.F.fq.gz\n", "Renaming ./2F_0_R1_.fastq.gz -> Pop2_SampleF.F.fq.gz\n", "Renaming ./3I_0_R1_.fastq.gz -> Pop3_SampleI.F.fq.gz\n", "Renaming ./1D_0_R1_.fastq.gz -> Pop1_SampleD.F.fq.gz\n", "export LD_LIBRARY_PATH=/home/iovercast/manuscript-analysis/dDocent//freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; export PATH=/home/iovercast/manuscript-analysis/dDocent/:$PATH; dDocent simlarge-config.txt\n", "dDocent 2.24 \n", "\n", "Contact jpuritz@gmail.com with any problems \n", "\n", " \n", "Checking for required software\n", "\n", "All required software is installed!\n", "dDocent will require input during the assembly stage. Please wait until prompt says it is safe to move program to the background.\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "Removing the _1 character and replacing with /1 in the name of every sequence\n", "Trimming reads and simultaneously assembling reference sequences\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n", "\n", "gzip: stdout: Broken pipe\n" ] } ], "source": [ "import ipyrad as ip\n", "import subprocess\n", "import glob\n", "\n", "## Set up directory structures. change the force flag if you want to\n", "## blow everything away and restart\n", "# force = True\n", "force = \"\"if force and os.path.exists(DDOCENT_OUTPUT):\n", " shutil.rmtree(DDOCENT_OUTPUT)\n", "if not os.path.exists(DDOCENT_OUTPUT):\n", " os.makedirs(DDOCENT_OUTPUT)\n", "os.chdir(DDOCENT_OUTPUT)\n", "\n", "## Have to copy all the raw files here so ddocent can find them\n", "IPYRAD_EDITS_DIR = os.path.join(IPYRAD_OUTPUT, \"REALDATA_edits/\")\n", "infiles = [ff for ff in glob.glob(IPYRAD_EDITS_DIR+\"*_R1_*\")]\n", "## Don't recopy the data over and over again while testing\n", "if not os.path.exists(infiles[0].split(\"/\")[-1]):\n", " for f in infiles:\n", " shutil.copy2(f, f.split(\"/\")[-1])\n", "\n", "## Now we have to rename all the files in the way dDocent expects them:\n", "## 1A_0_R1_.fastq.gz -> Pop1_Sample1.F.fq.gz\n", "name_mapping = {\"29154_superba_R1_.fastq\":\"Pop1_Sample1.F.fq\", \\\n", " \"32082_przewalskii_R1_.fastq\":\"Pop2_Sample1.F.fq\", \\\n", " \"33588_przewalskii_R1_.fastq\":\"Pop2_Sample2.F.fq\",\\\n", " \"35236_rex_R1_.fastq\":\"Pop3_Sample1.F.fq\", \"39618_rex_R1_.fastq\":\"Pop3_Sample2.F.fq\", \\\n", " \"35855_rex_R1_.fastq\":\"Pop3_Sample3.F.fq\", \"40578_rex_R1_.fastq\":\"Pop3_Sample4.F.fq\", \\\n", " \"38362_rex_R1_.fastq\":\"Pop3_Sample5.F.fq\", \\\n", " \"41954_cyathophylloides_R1_.fastq\":\"Pop4_Sample1.F.fq\", \"30686_cyathophylla_R1_.fastq\":\"Pop4_Sample2.F.fq\", \\\n", " \"41478_cyathophylloides_R1_.fastq\":\"Pop4_Sample3.F.fq\",\\\n", " \"30556_thamno_R1_.fastq\":\"Pop5_Sample1.F.fq\", \"33413_thamno_R1_.fastq\":\"Pop5_Sample2.F.fq\"}\n", "for k,v in name_mapping.items():\n", " os.rename(k, v)\n", "\n", "## Only runs on gzip files.\n", "!gzip *.fq\n", "\n", "## Write out the config file for this run.\n", "## Compacted the config file into one long line here to make it not take up so much room\n", "config_file = \"empirical-config.txt\".format(dir)\n", "with open(config_file, 'w') as outfile:\n", " outfile.write('Number of Processors\\n40\\nMaximum Memory\\n0\\nTrimming\\nyes\\nAssembly?\\nyes\\nType_of_Assembly\\nSE\\nClustering_Similarity%\\n0.85\\nMapping_Reads?\\nyes\\nMapping_Match_Value\\n1\\nMapping_MisMatch_Value\\n3\\nMapping_GapOpen_Penalty\\n5\\nCalling_SNPs?\\nyes\\nEmail\\nwatdo@mailinator.com\\n')\n", "\n", "cmd = \"export LD_LIBRARY_PATH={}/freebayes-src/vcflib/tabixpp/htslib/:$LD_LIBRARY_PATH; \".format(DDOCENT_DIR)\n", "cmd += \"export PATH={}:$PATH; time dDocent {}\".format(DDOCENT_DIR, config_file)\n", "print(cmd)\n", "## Have to run the printed command by hand from the ddocent REALDATA dir bcz it doesn't like running in the notebook\n", "#!$cmd\n", "\n", "## NB: Must rename all the samples in the output vcf and then use vcf-shuffle-cols\n", "## perl script in the vcf/perl directory to reorder the vcf file to match\n", "## the output of stacks and ipyrad for pca/heatmaps to work.\n" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "dDocent makes you rename your samples and then it orders the vcf according to \n", "these new sample names, so the order is different from ipyrad/pyrad/stacks\n", "which fsck plotting downstream, so we have to relabel all the columns\n", "to have the familiar names and then reorder them to be in the same order as the other progs." ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": true }, "outputs": [], "source": [ "## There's probably a better way to do this\n", "for f in [\"Final.recode.vcf\", \"TotalRawSNPs.vcf\"]:\n", " vcffile = os.path.join(DDOCENT_OUTPUT, f)\n", " infile = open(vcffile,'r')\n", " filedata = infile.readlines()\n", " infile.close()\n", " \n", " outfile = open(vcffile,'w')\n", " for line in filedata:\n", " if \"CHROM\" in line:\n", " for ipname, ddname in name_mapping.items():\n", " ipname = ipname.split(\"_R1\")[0]\n", " ddname = ddname.split(\".\")[0]\n", " line = line.replace(ddname, ipname)\n", " outfile.write(line)\n", " outfile.close()\n", " \n", " ## Now we have to reorder the genotype columns\n", " IPYRAD_VCF = os.path.join(IPYRAD_OUTPUT, \"REALDATA_outfiles/REALDATA.vcf\")\n", " os.chdir(os.path.join(DDOCENT_DIR, \"vcftools_0.1.11/perl\"))\n", " tmpvcf = os.path.join(DDOCENT_DIR, \"ddocent-tmp.vcf\")\n", " cmd = \"perl vcf-shuffle-cols -t {} {} > {}\".format(IPYRAD_VCF, vcffile, tmpvcf)\n", " print(cmd)\n", " os.system(cmd)\n", " os.rename(tmpvcf, vcffile)" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "# Results" ] }, { "cell_type": "markdown", "metadata": { "collapsed": true }, "source": [ "\n", "## Addendum\n", "\n", "This is me carping about organization of the stacks manual. The walkthrough doesn't mention\n", "that you _don't_ need to use mysql until somewhere in the late-middle of the process, long\n", "after you actually have to do the install in order to get it running.\n", "```\n", "## NB: If you pass the `-S` flag to denovo_map.pl then it won't require the sql backend. \n", "## This message is totally buried in the docs.\n", "\n", "## Also `--overw_db` and `--create_db` flags would have been good to know about.\n", "\n", "## denovo_map.pl refused to run w/o mysql (command not found error) so I had to brew install it\n", "brew install mysql\n", "mysql.server start\n", "msyql -u root\n", "> GRANT ALL ON *.* TO 'stacks_user'@'localhost' IDENTIFIED BY 'stackspassword';\n", "> create database stacks\n", "\n", "## Populate the mysql database with the right tables\n", "mysql -u root stacks < /usr/local/share/stacks/sql/stacks.sql\n", "\n", "## Edit the /usr/local/share/stacks/sql/mysql.cnf.dist file to reflect the username and password as above\n", "## then copy this file to /usr/local/share/stacks/sql/mysql.cnf\n", "```\n" ] }, { "cell_type": "code", "execution_count": null, "metadata": { "collapsed": true }, "outputs": [], "source": [] } ], "metadata": { "anaconda-cloud": {}, "kernelspec": { "display_name": "Python [default]", "language": "python", "name": "python2" }, "language_info": { "codemirror_mode": { "name": "ipython", "version": 2 }, "file_extension": ".py", "mimetype": "text/x-python", "name": "python", "nbconvert_exporter": "python", "pygments_lexer": "ipython2", "version": "2.7.11" } }, "nbformat": 4, "nbformat_minor": 0 }