{ "cells": [ { "cell_type": "markdown", "metadata": {}, "source": [ "# Indexing Variaton" ] }, { "cell_type": "code", "execution_count": 1, "metadata": {}, "outputs": [ { "data": { "text/html": [ "" ], "text/vnd.plotly.v1+html": [ "" ] }, "metadata": {}, "output_type": "display_data" } ], "source": [ "import scipy\n", "import numpy as np\n", "from sklearn.neighbors import KernelDensity\n", "from sklearn.decomposition import PCA\n", "from sklearn.model_selection import GridSearchCV\n", "from sklearn.cluster import estimate_bandwidth\n", "from sklearn.cluster import MeanShift, estimate_bandwidth\n", "\n", "import pandas as pd\n", "\n", "from scipy import stats\n", "from scipy.stats import beta\n", "from math import sin\n", "from random import randint\n", "\n", "###########################################\n", "import matplotlib.pyplot as plt\n", "import matplotlib\n", "\n", "%matplotlib inline\n", "\n", "import itertools as it\n", "\n", "\n", "\n", "from matplotlib.collections import BrokenBarHCollection\n", "import pandas as pd\n", "\n", "import re\n", "\n", "import plotly\n", "import plotly.plotly as py\n", "import plotly.graph_objs as go\n", "from plotly.offline import download_plotlyjs, init_notebook_mode, plot, iplot\n", "from plotly.graph_objs import *\n", "\n", "from ipywidgets import interact, interactive, fixed, interact_manual\n", "import ipywidgets as widgets\n", "init_notebook_mode(connected=True)" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "In previous posts, we resorted to density estimates to attempt to weed out outliers, identify cases of mislabelling and classify unlabelled observations. \n", "\n", "We went on to explore how cluster-derived KDE vectors of the same variables would aggregate in feature space, allowing us to label them in bulk (see **5. Visualizing KDE**).\n", "\n", "Varying distances among clusters, resulting distinguishable cluster profiles, can be of interest, depending on the nature of the data we are considering. If those distances vary according to physical distance for example, it might be interesting to extract and visualize those locations.\n", "\n", "Let's begin by performing the same simulations, of three populations, of which two see their distributions overlap as a function of a variable X (also seen in **6. Conditional Variation**)." ] }, { "cell_type": "code", "execution_count": 2, "metadata": {}, "outputs": [], "source": [ "\n", "# We must first define the number of populations, the length of the haplotypes desired, and their respective population sizes\n", "L= 300\n", "\n", "import itertools as it\n", "n= 20\n", "n_samples= 20\n", "\n", "a_range= np.linspace(1.5,2.5,n_samples)\n", "a_set= [i for i in a_range for _ in range(n)]\n", "\n", "b_range= np.linspace(0.1,.6,n_samples)\n", "b_set= [i for i in b_range for _ in range(n)]\n", "\n", "L_set= [L] * n * n_samples\n", "\n", "traces= [x for x in it.product(range(n*11),range(n*11),range(n*11))]\n", "\n", "background= np.array([a_set,b_set,L_set]).T\n", "\n", "vector_lib= []\n", "for k in range(background.shape[0]):\n", " \n", " probs= beta.rvs(background[k,0], background[k,1], size=int(background[k,2]))\n", " probs[(probs > 1)]= 1\n", " probs= [1-x for x in probs]\n", " \n", " \n", " vector_lib.append(probs)\n", "\n", "vector_lib= np.array(vector_lib)\n", "\n" ] }, { "cell_type": "code", "execution_count": 3, "metadata": {}, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "PC1: 0.047; PC2: 0.011; PC3: 0.011; PC4: 0.011; PC5: 0.01; PC6: 0.01; PC7: 0.01; PC8: 0.01; PC9: 0.01; PC10: 0.01; PC11: 0.009; PC12: 0.009; PC13: 0.009; PC14: 0.009; PC15: 0.009; PC16: 0.009; PC17: 0.009; PC18: 0.009; PC19: 0.009; PC20: 0.009; PC21: 0.008; PC22: 0.008; PC23: 0.008; PC24: 0.008; PC25: 0.008; PC26: 0.008; PC27: 0.008; PC28: 0.008; PC29: 0.008; PC30: 0.008; PC31: 0.008; PC32: 0.007; PC33: 0.007; PC34: 0.007; PC35: 0.007; PC36: 0.007; PC37: 0.007; PC38: 0.007; PC39: 0.007; PC40: 0.007; PC41: 0.007; PC42: 0.007; PC43: 0.007; PC44: 0.006; PC45: 0.006; PC46: 0.006; PC47: 0.006; PC48: 0.006; PC49: 0.006; PC50: 0.006\n", "features shape: (400, 50)\n" ] } ], "source": [ "n_comp = 50\n", "\n", "pca = PCA(n_components=n_comp, whiten=False,svd_solver='randomized')\n", "features = pca.fit_transform(vector_lib)\n", "\n", "var_comps= pca.explained_variance_ratio_\n", "print(\"; \".join(['PC{0}: {1}'.format(x+1,round(var_comps[x],3)) for x in range(n_comp)]))\n", "print('features shape: {}'.format(features.shape))" ] }, { "cell_type": "code", "execution_count": 4, "metadata": {}, "outputs": [ { "data": { "application/vnd.plotly.v1+json": { "data": [ { "marker": { "line": { "width": 0 }, "opacity": 0.8, "size": 4, "symbol": "circle" }, "mode": "markers", "text": [ "a: 1.5; b: 0.1, L: 300.0; index = 0", "a: 1.5; b: 0.1, L: 300.0; index = 1", "a: 1.5; b: 0.1, L: 300.0; index = 2", "a: 1.5; b: 0.1, L: 300.0; index = 3", "a: 1.5; b: 0.1, L: 300.0; index = 4", "a: 1.5; b: 0.1, L: 300.0; index = 5", "a: 1.5; b: 0.1, L: 300.0; index = 6", "a: 1.5; b: 0.1, L: 300.0; index = 7", "a: 1.5; b: 0.1, L: 300.0; index = 8", "a: 1.5; b: 0.1, L: 300.0; index = 9", "a: 1.5; b: 0.1, L: 300.0; index = 10", "a: 1.5; b: 0.1, L: 300.0; index = 11", "a: 1.5; b: 0.1, L: 300.0; index = 12", "a: 1.5; b: 0.1, L: 300.0; index = 13", "a: 1.5; b: 0.1, L: 300.0; index = 14", "a: 1.5; b: 0.1, L: 300.0; index = 15", "a: 1.5; b: 0.1, L: 300.0; index = 16", "a: 1.5; b: 0.1, L: 300.0; index = 17", "a: 1.5; b: 0.1, L: 300.0; index = 18", "a: 1.5; b: 0.1, L: 300.0; index = 19", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 20", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 21", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 22", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 23", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 24", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 25", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 26", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 27", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 28", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 29", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 30", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 31", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 32", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 33", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 34", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 35", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 36", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 37", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 38", "a: 1.5526315789473684; b: 0.12631578947368421, L: 300.0; index = 39", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 40", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 41", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 42", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 43", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 44", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 45", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 46", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 47", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 48", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 49", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 50", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 51", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 52", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 53", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 54", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 55", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 56", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 57", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 58", "a: 1.6052631578947367; b: 0.15263157894736842, L: 300.0; index = 59", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 60", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 61", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 62", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 63", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 64", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 65", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 66", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 67", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 68", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 69", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 70", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 71", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 72", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 73", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 74", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 75", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 76", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 77", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 78", "a: 1.6578947368421053; b: 0.17894736842105263, L: 300.0; index = 79", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 80", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 81", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 82", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 83", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 84", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 85", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 86", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 87", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 88", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 89", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 90", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 91", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 92", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 93", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 94", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 95", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 96", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 97", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 98", "a: 1.7105263157894737; b: 0.20526315789473684, L: 300.0; index = 99", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 100", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 101", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 102", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 103", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 104", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 105", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 106", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 107", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 108", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 109", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 110", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 111", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 112", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 113", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 114", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 115", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 116", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 117", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 118", "a: 1.763157894736842; b: 0.23157894736842105, L: 300.0; index = 119", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 120", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 121", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 122", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 123", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 124", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 125", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 126", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 127", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 128", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 129", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 130", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 131", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 132", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 133", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 134", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 135", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 136", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 137", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 138", "a: 1.8157894736842106; b: 0.2578947368421053, L: 300.0; index = 139", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 140", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 141", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 142", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 143", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 144", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 145", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 146", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 147", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 148", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 149", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 150", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 151", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 152", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 153", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 154", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 155", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 156", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 157", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 158", "a: 1.868421052631579; b: 0.28421052631578947, L: 300.0; index = 159", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 160", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 161", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 162", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 163", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 164", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 165", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 166", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 167", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 168", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 169", "a: 1.9210526315789473; b: 0.31052631578947365, L: 300.0; index = 170", 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310", "a: 2.2894736842105265; b: 0.49473684210526314, L: 300.0; index = 311", "a: 2.2894736842105265; b: 0.49473684210526314, L: 300.0; index = 312", "a: 2.2894736842105265; b: 0.49473684210526314, L: 300.0; index = 313", "a: 2.2894736842105265; b: 0.49473684210526314, L: 300.0; index = 314", "a: 2.2894736842105265; b: 0.49473684210526314, L: 300.0; index = 315", "a: 2.2894736842105265; b: 0.49473684210526314, L: 300.0; index = 316", "a: 2.2894736842105265; b: 0.49473684210526314, L: 300.0; index = 317", "a: 2.2894736842105265; b: 0.49473684210526314, L: 300.0; index = 318", "a: 2.2894736842105265; b: 0.49473684210526314, L: 300.0; index = 319", "a: 2.3421052631578947; b: 0.5210526315789473, L: 300.0; index = 320", "a: 2.3421052631578947; b: 0.5210526315789473, L: 300.0; index = 321", "a: 2.3421052631578947; b: 0.5210526315789473, L: 300.0; index = 322", "a: 2.3421052631578947; b: 0.5210526315789473, L: 300.0; index = 323", "a: 2.3421052631578947; b: 0.5210526315789473, L: 300.0; index 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" ] }, "metadata": {}, "output_type": "display_data" } ], "source": [ "#### Let's plot the first 3 coordinates nonetheless.\n", "####\n", "fig_data= [go.Scatter3d(\n", " x = features[:,0],\n", " y = features[:,1],\n", " z = features[:,2],\n", " type='scatter3d',\n", " mode= \"markers\",\n", " text= ['a: {}; b: {}, L: {}; index = {}'.format(background[k,0],background[k,1],background[k,2], k) for k in range(background.shape[0])],\n", " marker= {\n", " 'line': {'width': 0},\n", " 'size': 4,\n", " 'symbol': 'circle',\n", " \"opacity\": .8\n", " }\n", " )]\n", "\n", "\n", "layout = go.Layout(\n", " margin=dict(\n", " l=0,\n", " r=0,\n", " b=0,\n", " t=0\n", " ),\n", " scene= Scene(\n", " yaxis=dict(\n", " title='PC2: {}'.format(round(var_comps[1],3))),\n", " xaxis=dict(\n", " title= 'PC1: {}'.format(round(var_comps[0],3))),\n", " zaxis=dict(\n", " title= 'PC3: {}'.format(round(var_comps[2],3))))\n", ")\n", "\n", "fig = go.Figure(data=fig_data, layout=layout)\n", "iplot(fig)\n" ] }, { "cell_type": "code", "execution_count": 5, "metadata": {}, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "global fst: 0.1513\n" ] }, { "data": { "text/html": [ "
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2(353, 375)0.093620
\n", "
" ], "text/plain": [ " pops fst\n", "0 (363, 353) 0.112383\n", "1 (363, 375) 0.109360\n", "2 (353, 375) 0.093620" ] }, "execution_count": 5, "metadata": {}, "output_type": "execute_result" } ], "source": [ "#### calculating Fsts between populations.\n", "Pops= [363,353,375]\n", "Sizes= [80,100,30]\n", "\n", "def return_fsts(vector_lib,pops):\n", " H= {pop: [1-(vector_lib[pop,x]**2 + (1 - vector_lib[pop,x])**2) for x in range(vector_lib.shape[1])] for pop in pops}\n", " Store= []\n", " for comb in it.combinations(pops,2):\n", " P= [sum([vector_lib[x,i] for x in comb]) / len(comb) for i in range(vector_lib.shape[1])]\n", " HT= [2 * P[x] * (1 - P[x]) for x in range(len(P))]\n", " Fst= np.mean([(HT[x] - np.mean([H[p][x] for p in comb])) / HT[x] for x in range(len(P))])\n", " \n", " Store.append([comb,Fst])\n", " \n", " ### total fst:\n", " P= [sum([vector_lib[x,i] for x in pops]) / len(pops) for i in range(vector_lib.shape[1])]\n", " HT= [2 * P[x] * (1 - P[x]) for x in range(len(P))]\n", " FST= np.mean([(HT[x] - np.mean([H[p][x] for p in pops])) / HT[x] for x in range(len(P))])\n", " \n", " return pd.DataFrame(Store,columns= ['pops','fst']),FST\n", "\n", "Pairwise, Total_fst= return_fsts(vector_lib,Pops)\n", "\n", "print('global fst: {}'.format(round(Total_fst,4)))\n", "Pairwise" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "We'll now go through the same motions, so that we can attest, through visual analysis, which groups of vectors represent which kinds of clusters." ] }, { "cell_type": "code", "execution_count": 6, "metadata": {}, "outputs": [], "source": [ "\n", "def extract_profiles(global_data):\n", " ## estimate the bandwith\n", " bandwidth = estimate_bandwidth(global_data, quantile=0.2)\n", " params = {'bandwidth': np.linspace(np.min(global_data), np.max(global_data),20)}\n", " grid = GridSearchCV(KernelDensity(algorithm = \"ball_tree\",breadth_first = False), params,verbose=0)\n", " \n", " ## perform MeanShift clustering.\n", " ms = MeanShift(bandwidth=bandwidth, bin_seeding=True, cluster_all=True, min_bin_freq=5)\n", " ms.fit(global_data)\n", " labels1 = ms.labels_\n", " label_select = {y:[x for x in range(len(labels1)) if labels1[x] == y] for y in sorted(list(set(labels1)))}\n", "\n", " ## Extract the KDE of each cluster identified by MS.\n", " cluster_profiles= []\n", "\n", " \n", "\n", " for lab in label_select.keys():\n", " \n", " Quanted_set= global_data[label_select[lab],:]\n", "\n", " grid.fit(Quanted_set)\n", "\n", " kde = grid.best_estimator_\n", "\n", " P_dist = kde.score_samples(Quanted_set)\n", " Fist = kde.score_samples(global_data)\n", "\n", " ## Normalizing log-likelihood estimates by those of the reference set.\n", " Fist = scipy.stats.norm(np.mean(P_dist),np.std(P_dist)).cdf(Fist)\n", " cluster_profiles.append(Fist)\n", " \n", " return cluster_profiles\n", " \n", "#### We'll have to first change our fst function.\n", "\n", "def return_fsts2(freq_array):\n", " pops= range(freq_array.shape[0])\n", " H= {pop: [1-(freq_array[pop,x]**2 + (1 - freq_array[pop,x])**2) for x in range(freq_array.shape[1])] for pop in range(freq_array.shape[0])}\n", " Store= []\n", "\n", " for comb in it.combinations(H.keys(),2):\n", " P= [sum([freq_array[x,i] for x in comb]) / len(comb) for i in range(freq_array.shape[1])]\n", " HT= [2 * P[x] * (1 - P[x]) for x in range(len(P))]\n", " per_locus_fst= [[(HT[x] - np.mean([H[p][x] for p in comb])) / HT[x],0][int(HT[x] == 0)] for x in range(len(P))]\n", " per_locus_fst= np.nan_to_num(per_locus_fst)\n", " Fst= np.mean(per_locus_fst)\n", "\n", " Store.append([comb,Fst])\n", " \n", " \n", " ### total fst:\n", " P= [sum([freq_array[x,i] for x in pops]) / len(pops) for i in range(freq_array.shape[1])]\n", " HT= [2 * P[x] * (1 - P[x]) for x in range(len(P))]\n", " FST= np.mean([(HT[x] - np.mean([H[p][x] for p in pops])) / HT[x] for x in range(len(P))])\n", " \n", " return pd.DataFrame(Store,columns= ['pops','fst'])\n" ] }, { "cell_type": "code", "execution_count": 7, "metadata": {}, "outputs": [ { "name": "stderr", "output_type": "stream", "text": [ "c:\\users\\jgarcia\\appdata\\local\\programs\\python\\python36-32\\lib\\site-packages\\ipykernel_launcher.py:46: RuntimeWarning:\n", "\n", "invalid value encountered in double_scalars\n", "\n" ] } ], "source": [ "#### We're going to do something different now. We'll have two points get closer together in time.\n", "#first chose two\n", "target= [1,2]\n", "\n", "\n", "labels= []\n", "Fsts_crawl= []\n", "angle_list= []\n", "Distances_crawl= []\n", "\n", "for angle in np.arange(1,30,.1):\n", " coords= features[Pops,:]\n", " vector2= coords[target[1]] - coords[target[0]]\n", "\n", " coords[target[0]] = coords[target[0]] + [sin(angle) * x for x in vector2]\n", " \n", " new_freqs= pca.inverse_transform(coords)\n", " \n", " new_freqs[target[0]]= pca.inverse_transform(coords[target[0]])\n", " new_freqs[new_freqs > 1] = 1\n", " new_freqs[new_freqs < 0] = 0\n", " \n", " Pairwise= return_fsts2(new_freqs)\n", " \n", " Distances= []\n", " for train in it.combinations([x for x in range(new_freqs.shape[0])],2):\n", " Distances.append(np.sqrt((coords[train[0]][0] - coords[train[1]][0])**2 + (coords[train[0]][1] - coords[train[1]][1])**2) + (coords[train[0]][2] - coords[train[1]][2])**2)\n", " Distances_crawl.extend(Distances)\n", " \n", " labels.extend(Pairwise.pops)\n", " \n", " Fsts_crawl.extend(Pairwise.fst)\n", " angle_list.extend([angle] * Pairwise.shape[0])\n", "\n", "\n", "Control= np.array([angle_list,Fsts_crawl]).T" ] }, { "cell_type": "code", "execution_count": 8, "metadata": {}, "outputs": [ { "data": { "application/vnd.plotly.v1+json": { "data": [ { "mode": "markers", "name": "fst vs distances", "type": "scatter", "x": [ 0.7045945662144438, 0.708741321836562, 0.02960043355665976, 0.7057465485888003, 0.708741321836562, 0.02031063050489924, 0.7067945622911205, 0.708741321836562, 0.012686136681670096, 0.7076661386985985, 0.708741321836562, 0.006801875844128669, 0.708303381569976, 0.708741321836562, 0.0027156322317604597, 0.7086651076328876, 0.708741321836562, 0.0004675134449482361, 0.7087283248685391, 0.708741321836562, 7.957843891132374e-05, 0.708489042069622, 0.708741321836562, 0.0015556332043705665, 0.7079624082346324, 0.708741321836562, 0.004881195535114318, 0.7071821817013597, 0.708741321836562, 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" ], "text/vnd.plotly.v1+html": [ "
" ] }, "metadata": {}, "output_type": "display_data" } ], "source": [ "coords= {z:[x for x in range(len(labels)) if labels[x] == z] for z in list(set(labels))}\n", "\n", "fig_data= [go.Scatter(\n", "x= [sin(x) for x in Control[coords[i],0]],\n", "y= Control[coords[i],1],\n", "mode= 'lines',\n", "name= 'fst pops {} / {}'.format(i[0],i[1])\n", ") for i in coords.keys()\n", "]\n", "\n", "layout = go.Layout(\n", " title= 'pairwise Fsts against the sinusoid of variable X',\n", " yaxis=dict(\n", " title='between population Fst'),\n", " xaxis=dict(\n", " title='sin(X)')\n", ")\n", "\n", "fig= go.Figure(data=fig_data, layout=layout)\n", "iplot(fig)" ] }, { "cell_type": "code", "execution_count": 11, "metadata": {}, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "Variance explained:\n", "PC1: 0.491; PC2: 0.16; PC3: 0.023\n" ] } ], "source": [ "labels= [0,1,2]\n", "\n", "label_vector= np.repeat(np.array([x for x in labels]),Sizes)\n", "label_indicies= {x:[y for y in range(len(label_vector)) if label_vector[y] == x] for x in labels}\n", "\n", "cluster_profiles= []\n", "\n", "Windows= []\n", "\n", "\n", "target_indx= {z:[x for x in range(len(label_vector)) if label_vector[x] == z] for z in list(set(label_vector))}\n", "threshold= .005\n", "P= 30\n", "\n", "\n", "for angle in np.arange(1,30,.1):\n", " coords= features[Pops,:]\n", " vector2= coords[target[1]] - coords[target[0]]\n", "\n", " coords[target[0]] = coords[target[0]] + [sin(angle) * x for x in vector2]\n", " \n", " new_freqs= pca.inverse_transform(coords)\n", " #new_freqs= vector_lib[Pops,:]\n", " new_freqs[target[0]]= pca.inverse_transform(coords[target[0]])\n", "\n", "\n", " N_pops= len(Pops)\n", " \n", " labels= np.repeat(np.array([x for x in range(N_pops)]),Sizes)\n", "\n", " data= []\n", "\n", " for k in range(N_pops):\n", "\n", " probs= new_freqs[k,:]\n", " \n", " probs[(probs > 1)]= 1\n", " probs[(probs < 0)]= 0\n", " m= Sizes[k]\n", " Haps= [[np.random.choice([1,0],p= [1-probs[x],probs[x]]) for x in range(L)] for acc in range(m)]\n", "\n", " data.extend(Haps)\n", " \n", " data= np.array(data)\n", " pca2 = PCA(n_components=3, whiten=False,svd_solver='randomized')\n", "\n", " data= pca2.fit_transform(data)\n", "\n", " profiles= extract_profiles(data,)\n", " \n", " ### store stuff.\n", " cluster_profiles.extend(profiles)\n", " Windows.extend([angle] * len(profiles))\n", "\n", "\n", "## number of comps to retain\n", "n_comp= 3\n", "\n", "pca = PCA(n_components=n_comp, whiten=False).fit(cluster_profiles)\n", "COMPS = pca.transform(cluster_profiles)\n", "X_se = pca.components_.T*np.sqrt(pca.explained_variance_)\n", "\n", "print(\"Variance explained:\")\n", "print(\"; \".join(['PC{0}: {1}'.format(x+1,round(pca.explained_variance_ratio_[x],3)) for x in range(n_comp)]))## stacking our data.\n" ] }, { "cell_type": "code", "execution_count": 13, "metadata": {}, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "(210, 3)\n", "(742, 3)\n" ] }, 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" ] }, "metadata": {}, "output_type": "display_data" } ], "source": [ "print(X_se.shape)\n", "print(COMPS.shape)\n", "\n", "fig_data= [go.Scatter3d(\n", " x = X_se[[x for x in range(sum(Sizes)) if label_vector[x] == i],0],\n", " y = X_se[[x for x in range(sum(Sizes)) if label_vector[x] == i],1],\n", " z = X_se[[x for x in range(sum(Sizes)) if label_vector[x] == i],2],\n", " type='scatter3d',\n", " mode= \"markers\",\n", " marker= {\n", " 'line': {'width': 0},\n", " 'size': 4,\n", " 'symbol': 'circle',\n", " \"opacity\": .8\n", " },\n", " name= str(i)\n", " ) for i in list(set(labels))]\n", "\n", "\n", "\n", "layout = go.Layout(\n", " margin=dict(\n", " l=0,\n", " r=0,\n", " b=0,\n", " t=0\n", " )\n", ")\n", "fig = go.Figure(data=fig_data, layout=layout)\n", "iplot(fig)\n" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "Relative distance of observations given the KDE profiles extracted." ] }, { "cell_type": "code", "execution_count": 15, "metadata": { "scrolled": false }, 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" ], "text/vnd.plotly.v1+html": [ "
" ] }, "metadata": {}, "output_type": "display_data" } ], "source": [ "bandwidth = estimate_bandwidth(COMPS, quantile=0.2)\n", "params = {'bandwidth': np.linspace(np.min(COMPS), np.max(COMPS),20)}\n", "grid = GridSearchCV(KernelDensity(algorithm = \"ball_tree\",breadth_first = False), params,verbose=0)\n", "\n", "## perform MeanShift clustering.\n", "ms = MeanShift(bandwidth=bandwidth, bin_seeding=True, cluster_all=True, min_bin_freq=5)\n", "ms.fit(COMPS)\n", "labels1 = ms.labels_\n", "\n", "label_select = {y:[x for x in range(len(labels1)) if labels1[x] == y] for y in sorted(list(set(labels1)))}\n", "\n", "fig_data= [go.Scatter3d(\n", " x = COMPS[label_select[i],0],\n", " y = COMPS[label_select[i],1],\n", " z = COMPS[label_select[i],2],\n", " type='scatter3d',\n", " mode= \"markers\",\n", " marker= {\n", " 'line': {'width': 0},\n", " 'size': 4,\n", " 'symbol': 'circle',\n", " \"opacity\": .8\n", " },\n", " name= str(i)\n", " ) for i in label_select.keys()]\n", "\n", "\n", "\n", "layout = go.Layout(\n", " margin=dict(\n", " l=0,\n", " r=0,\n", " b=0,\n", " t=0\n", " )\n", ")\n", "fig = go.Figure(data=fig_data, layout=layout)\n", "iplot(fig)\n" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "KDE profiles in feature space. Colors represent MeanShift identified clusters in this space." ] }, { "cell_type": "code", "execution_count": 16, "metadata": { "scrolled": false }, "outputs": [ { "data": { "application/vnd.jupyter.widget-view+json": { "model_id": "98d6366b71e24fc6b5e6fc337da32f4b", "version_major": 2, "version_minor": 0 }, "text/html": [ "

Failed to display Jupyter Widget of type interactive.

\n", "

\n", " If you're reading this message in the Jupyter Notebook or JupyterLab Notebook, it may mean\n", " that the widgets JavaScript is still loading. If this message persists, it\n", " likely means that the widgets JavaScript library is either not installed or\n", " not enabled. See the Jupyter\n", " Widgets Documentation for setup instructions.\n", "

\n", "

\n", " If you're reading this message in another frontend (for example, a static\n", " rendering on GitHub or NBViewer),\n", " it may mean that your frontend doesn't currently support widgets.\n", "

\n" ], "text/plain": [ "interactive(children=(Dropdown(description='pop', options=('labels', '0', '1', '2', '3', '4'), value='labels'), Output()), _dom_classes=('widget-interact',))" ] }, "metadata": {}, "output_type": "display_data" }, { "data": { "text/plain": [ "" ] }, "execution_count": 16, "metadata": {}, "output_type": "execute_result" } ], "source": [ "## Now let's grab those labels and see who those clusters represent:\n", "cluster_profiles= np.array(cluster_profiles)\n", "Score= {i:np.mean(cluster_profiles[label_select[i],:],axis= 0) for i in label_select.keys()}\n", "\n", "\n", "Titles= ['labels']\n", "Titles.extend([str(x) for x in label_select.keys()])\n", "\n", "def figure_scores(pop):\n", " \n", " if pop not in ['labels']:\n", " fig_data= [go.Scatter3d(\n", " x = X_se[:,0],\n", " y = X_se[:,1],\n", " z = X_se[:,2],\n", " type='scatter3d',\n", " mode= \"markers\",\n", " marker= {\n", " 'color': Score[int(pop)],\n", " 'colorbar': go.ColorBar(\n", " title= 'ColorBar'\n", " ),\n", " 'colorscale': 'Viridis',\n", " 'line': {'width': 0},\n", " 'size': 4,\n", " 'symbol': 'circle',\n", " \"opacity\": .8\n", " }\n", " )]\n", " else:\n", " coords= {y:[x for x in range(len(label_vector)) if label_vector[x] == y] for y in labels}\n", " fig_data= [go.Scatter3d(\n", " x = X_se[coords[i],0],\n", " y = X_se[coords[i],1],\n", " z = X_se[coords[i],2],\n", " type='scatter3d',\n", " mode= \"markers\",\n", " marker= {\n", " 'line': {'width': 0},\n", " 'size': 4,\n", " 'symbol': 'circle',\n", " \"opacity\": .8\n", " }\n", " ) for i in coords.keys()]\n", " layout = go.Layout(\n", " margin=dict(\n", " l=0,\n", " r=0,\n", " b=0,\n", " t=0\n", " ),\n", " title= str(pop)\n", " )\n", " fig = go.Figure(data=fig_data, layout=layout)\n", " iplot(fig)\n", "\n", "interact(figure_scores, pop=Titles)" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "Average likelihood profiles within clusters inform us of the populations they represented. At least one of the clusters identified holds instances of clusters that capture two populations at once (i.e. when sin(x) == 0). Can you tell which?\n", "\n", "Now, because we were thinking ahead, we indexed each cluster profile to the value of 'X'. Let's represent each of our profiles by:\n", "- The cluster they were assigned to in feature space.\n", "- The value 'X' at which they were derived.\n", "- The sinusoid of 'X'.\n", "\n", "We'll set our Y axis to represent their assignments, so that they don't overlap, and Z to be their height, sin(X)." ] }, { "cell_type": "code", "execution_count": 17, "metadata": {}, "outputs": [ { "data": { "text/plain": [ "742" ] }, "execution_count": 17, "metadata": {}, "output_type": "execute_result" } ], "source": [ "sum([len(x) for x in label_select.values()])" ] }, { "cell_type": "code", "execution_count": 19, "metadata": { "scrolled": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "dict_keys([0, 1, 2, 3, 4])\n" ] }, { "data": { "application/vnd.plotly.v1+json": { "data": [ { "marker": { "line": { "width": 0 }, "opacity": 1, "size": 8, "symbol": "circle" }, "mode": "markers", "type": "scatter3d", "x": [ 1, 1.1, 1.2000000000000002, 1.3000000000000003, 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} }, "text/html": [ "
" ], "text/vnd.plotly.v1+html": [ "
" ] }, "metadata": {}, "output_type": "display_data" } ], "source": [ "print(label_select.keys())\n", "\n", "fig_data= [go.Scatter3d(\n", " x = [Windows[x] for x in label_select[i]],\n", " y = [i for x in label_select[i]],\n", " z = [sin(Windows[x]) for x in label_select[i]],\n", " type='scatter3d',\n", " mode= \"markers\",\n", " marker= {\n", " 'line': {'width': 0},\n", " 'size': 8,\n", " 'symbol': 'circle',\n", " \"opacity\": 1\n", " }\n", " ) for i in label_select.keys()]\n", "\n", "layout = go.Layout(\n", " margin=dict(\n", " l=0,\n", " r=0,\n", " b=0,\n", " t=0\n", " ),\n", " title= 'where'\n", ")\n", "\n", "fig = go.Figure(data=fig_data, layout=layout)\n", "iplot(fig)\n" ] }, { "cell_type": "markdown", "metadata": { "scrolled": false }, "source": [ "Here we can see that some clusters, the same that gave positive average likelihoods to elements of two groups, appear only as sin(X) nears 0. \n", "\n", "While their sensitivity will depend on the bandwidth used to estimate Kernel standard deviations as well as the converging population's relative covariances, their use in extracting X's relative proximity to 0 is evident.\n", "\n", "We might prefer to visualize this scenario in terms of which samples belonged to which class of vector profiles at a given window.\n", "\n", "The best representation of in this scenario is that of Ideograms, commonly used to describe classifications along multiple individual genomes.\n", "\n", "To do this, we must first examine which profile - and associated class, each sample was most likely assigned to at each window. " ] }, { "cell_type": "code", "execution_count": 20, "metadata": {}, "outputs": [], "source": [ "###### Mapping classifications.\n", "\n", "Ideo= []\n", "Window_list= {z:[x for x in range(len(Windows)) if Windows[x] == z] for z in list(set(Windows))}\n", "chromosome_list= []\n", "\n", "color_ref= ['blue','orange','green','red','purple','silver','green','silver','red3','deepskyeblue','navy','chartreuse','darkorchid3','goldenrod2']\n", "\n", "for window in Window_list.keys():\n", " Wind= int(window*1000)\n", " classes= np.argmax(cluster_profiles[Window_list[window],:],axis= 0)\n", " windows_labels= [labels1[x] for x in Window_list[window]]\n", " actual_labels= [windows_labels[x] for x in classes]\n", " for k in range(cluster_profiles.shape[1]):\n", " Ideo.append(['Region_' + str(1).zfill(2) + '_Ind' + str(k),Wind,Wind + 99,color_ref[actual_labels[k]]])\n", "\n", "\n", "for k in range(cluster_profiles.shape[1]):\n", " chromosome_list.append('Region_' + str(1).zfill(2) + '_Ind' + str(k))\n", "\n", "Out= {1:{int((x*1000)):int((x*1000)) + 99 for x in Windows}}" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "The table holding this information consists of a matrix with n_rows= n_windows * n_samples. Because not all samples change classification with every increment of X, we will compress this data. The resulting plot will be less heavy to compute." ] }, { "cell_type": "code", "execution_count": 57, "metadata": {}, "outputs": [], "source": [ "\n", "# Here's the function that we'll call for each dataframe (once for chromosome\n", "# ideograms, once for genes). The rest of this script will be prepping data\n", "# for input to this function\n", "#\n", "def chromosome_collections(df, y_positions, height, **kwargs):\n", " \"\"\"\n", " Yields BrokenBarHCollection of features that can be added to an Axes\n", " object.\n", " Parameters\n", " ----------\n", " df : pandas.DataFrame\n", " Must at least have columns ['chrom', 'start', 'end', 'color']. If no\n", " column 'width', it will be calculated from start/end.\n", " y_positions : dict\n", " Keys are chromosomes, values are y-value at which to anchor the\n", " BrokenBarHCollection\n", " height : float\n", " Height of each BrokenBarHCollection\n", " Additional kwargs are passed to BrokenBarHCollection\n", " \"\"\"\n", " del_width = False\n", " if 'width' not in df.columns:\n", " del_width = True\n", " df['width'] = df['end'] - df['start']\n", " for chrom, group in df.groupby('chrom'):\n", " \n", " yrange = (y_positions[chrom], height)\n", " xranges = group[['start', 'width']].values\n", " yield BrokenBarHCollection(\n", " xranges, yrange, facecolors=group['colors'], **kwargs)\n", " if del_width:\n", " del df['width']\n", "\n", "\n", "def compress_ideo(df,chromosome_list):\n", " \n", " new_set = []\n", " \n", " for CHR in range(len(chromosome_list)):\n", " \n", " Chr = int(re.search('Region_(.+?)_',chromosome_list[CHR]).group(1))\n", " sub = df[df.chrom == chromosome_list[CHR]]\n", " Coordinates = sorted(sub.start)\n", " Size = sub.shape[0]\n", " start = min(df.start)\n", " First = sub.gieStain.iloc[0]\n", " for index in range(len(Coordinates)):\n", " row = sub[sub.start == Coordinates[index]]\n", " if index == 0:\n", " continue\n", " if index == (Size - 1):\n", " if row.gieStain.iloc[0] == First:\n", " new_set.append([chromosome_list[CHR],start,Out[Chr][max(df.start)],First])\n", " else:\n", " new_set.append([chromosome_list[CHR],start,Out[Chr][max(df.start)],First])\n", " First = row.gieStain.iloc[0]\n", " start = row.start.iloc[0]\n", " new_set.append([chromosome_list[CHR],start,Out[Chr][max(df.start)],First])\n", " else:\n", " if row.gieStain.iloc[0] == First:\n", " continue\n", " else:\n", " new_set.append([chromosome_list[CHR],start,row.start.iloc[0]-1,First])\n", " First = row.gieStain.iloc[0]\n", " start = row.start.iloc[0]\n", " \n", " new_set = pd.DataFrame(new_set,columns = ['chrom', 'start', 'end', 'gieStain'])\n", " return new_set\n", "\n", "\n", "\n", "\n", "#### begin by compressing assignments by individuals. Lightens the load of the following plot.\n", "ideo = pd.DataFrame(Ideo,columns = ['chrom', 'start', 'end', 'gieStain'])\n", "\n", "# Filter out chromosomes not in our list\n", "ideo = ideo[ideo.chrom.apply(lambda x: x in chromosome_list)]\n", "\n", "ideo = compress_ideo(ideo,chromosome_list)\n" ] }, { "cell_type": "code", "execution_count": 58, "metadata": { "scrolled": false }, "outputs": [ { "name": "stdout", "output_type": "stream", "text": [ "adding ideograms...\n" ] }, { "data": { "image/png": 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\n", "text/plain": [ "" ] }, "metadata": {}, "output_type": "display_data" } ], "source": [ "# Here's the function that we'll call for each dataframe (once for chromosome\n", "# ideograms, once for genes). The rest of this script will be prepping data\n", "# for input to this function\n", "#\n", "def return_ideogram(ideo):\n", " # Height of each ideogram\n", " chrom_height = 1\n", "\n", " # Spacing between consecutive ideograms\n", " chrom_spacing = 0\n", "\n", " # Height of the gene track. Should be smaller than `chrom_spacing` in order to\n", " # fit correctly\n", " gene_height = 0.0\n", "\n", " # Padding between the top of a gene track and its corresponding ideogram\n", " gene_padding = 0.0\n", "\n", "\n", " # Keep track of the y positions for ideograms and genes for each chromosome,\n", " # and the center of each ideogram (which is where we'll put the ytick labels)\n", " ybase = 0\n", " chrom_ybase = {}\n", " gene_ybase = {}\n", " chrom_centers = {}\n", "\n", " # Iterate in reverse so that items in the beginning of `chromosome_list` will\n", " # appear at the top of the plot\n", " for chrom in chromosome_list[::-1]:\n", " chrom_ybase[chrom] = ybase\n", " chrom_centers[chrom] = ybase + chrom_height / 2.\n", " gene_ybase[chrom] = ybase - gene_height - gene_padding\n", " ybase += chrom_height + chrom_spacing\n", "\n", "\n", "\n", " # Keep track of the y positions for ideograms and genes for each chromosome,\n", " # and the center of each ideogram (which is where we'll put the ytick labels)\n", " ybase = 0\n", " chrom_ybase = {}\n", " gene_ybase = {}\n", " chrom_centers = {}\n", "\n", " # Iterate in reverse so that items in the beginning of `chromosome_list` will\n", " # appear at the top of the plot\n", " for chrom in chromosome_list[::-1]:\n", " chrom_ybase[chrom] = ybase\n", " chrom_centers[chrom] = ybase + chrom_height / 2.\n", " gene_ybase[chrom] = ybase - gene_height - gene_padding\n", " ybase += chrom_height + chrom_spacing\n", " \n", "\n", " # Colors for different chromosome stains\n", " color_lookup = {\n", " 'red': [255, 0, 0],\n", " 'yellow': [255, 255, 0],\n", " 'blue': [0, 0, 255],\n", " 'orange': [255, 165, 0],\n", " 'green': [50, 205, 50],\n", " 'black': [0, 0, 0],\n", " 'purple': [128, 0, 128],\n", " 'silver': [211, 211, 211],\n", " }\n", "\n", " # Add a new column for colors\n", " \n", " ideo['colors'] = ideo['gieStain'].apply(lambda x: tuple([round(y / float(255),1) for y in color_lookup[x]]))\n", " # Add a new column for width\n", " ideo['width'] = ideo.end - ideo.start\n", "\n", " # Width, height (in inches)\n", " figsize = (10, 30)\n", "\n", " fig = plt.figure(figsize=figsize)\n", " ax = fig.add_subplot(111)\n", "\n", " # Now all we have to do is call our function for the ideogram data...\n", " print(\"adding ideograms...\")\n", " for collection in chromosome_collections(ideo, chrom_ybase, chrom_height, edgecolors=None, linewidths= 0):\n", " ax.add_collection(collection)\n", "\n", " # Axes tweaking\n", " ax.set_xticks([x for x in range(min(ideo.start),max(ideo.end),int(1000))])\n", " plt.xticks(fontsize = 5,rotation = 90)\n", " ax.tick_params(axis = 'x',pad = 10)\n", "\n", " ax.tick_params(axis='y', which='major', pad=30)\n", " ax.set_yticks([chrom_centers[i] for i in chromosome_list])\n", " ax.set_yticklabels(chromosome_list, fontsize = 5)\n", " ax.axis('tight')\n", " return fig\n", "\n", "\n", "fig= return_ideogram(ideo)\n" ] }, { "cell_type": "markdown", "metadata": {}, "source": [ "The resulting ideogram: samples classified according to the clusters they belonged to as X varied.\n", "\n", "## Application\n", "\n", "If we didn't know that the distances between two of the groups varied as a sinusoid of X, we might be interested in examining what exactly was going on with individuals assigned to profiles encompassing the two groups at those locations they were assigned to them.\n", "\n", "In other contexts, location and extent of these assignments alone could be of interest." ] }, { "cell_type": "code", "execution_count": null, "metadata": {}, "outputs": [], "source": [] } ], "metadata": { "kernelspec": { "display_name": "Python 3", "language": "python", "name": "python3" }, "language_info": { "codemirror_mode": { "name": "ipython", "version": 3 }, "file_extension": ".py", "mimetype": "text/x-python", "name": "python", "nbconvert_exporter": "python", "pygments_lexer": "ipython3", "version": "3.6.3" } }, "nbformat": 4, "nbformat_minor": 2 }