#!/usr/bin/sh ## Charge tools you need for analysis module load star/2.7.5a module load samtools/1.9 module load subread/1.6.1 module load multiqc/1.7 #indexation of genome for STAR !!!!!!! DON'T RUN THIS COMMAND !!!! mkdir TAIR10 srun -c 4 STAR --runMode genomeGenerate --genomeFastaFiles Arabidopsis_thaliana.TAIR10.dna.toplevel.fa --genomeDir TAIR10 --runThreadN 4 srun samtools faidx Arabidopsis_thaliana.TAIR10.dna.toplevel.fa # Copy fastq files in your home: Choose one WT and one KO mkdir data cp -r /shared/projects/form_2022_32/coursLinux/rna-seq/01-Bioinfo/data/WT1* data/ cp -r /shared/projects/form_2022_32/coursLinux/rna-seq/01-Bioinfo/data/KO1* data/ # create a directory to store the future results mkdir results/ #example of command with one sample: srun -c 4 STAR --genomeDir /shared/projects/form_2022_32/coursLinux/rna-seq/01-Bioinfo/TAIR10 --runThreadN 4 --readFilesCommand zcat --outFileNamePrefix results/WT1_ --readFilesIn data/WT1_R1.fastq.gz data/WT1_R2.fastq.gz --outSAMtype BAM SortedByCoordinate --alignIntronMax 1000 --alignMatesGapMax 10000 --sjdbGTFfile /shared/projects/form_2022_32/coursLinux/rna-seq/01-Bioinfo/TAIR10/Arabidopsis_thaliana.TAIR10.45.gtf srun samtools index results/WT1_Aligned.sortedByCoord.out.bam # TODO: complete the command of featureCount. Google is your friend !! srun -c 4 featureCounts -T 4 ... #copy commands and change them to run all analysis (5min by sample) #KO1 srun -c 4 STAR --genomeDir /shared/projects/form_2022_32/coursLinux/rna-seq/01-Bioinfo/TAIR10 --runThreadN 4 --readFilesCommand zcat --outFileNamePrefix results/KO1_ --readFilesIn data/KO1_R1.fastq.gz data/KO1_R2.fastq.gz --outSAMtype BAM SortedByCoordinate --alignIntronMax 1000 --alignMatesGapMax 10000 --sjdbGTFfile /shared/projects/form_2022_32/coursLinux/rna-seq/01-Bioinfo/TAIR10/Arabidopsis_thaliana.TAIR10.45.gtf srun samtools index results/KO1_Aligned.sortedByCoord.out.bam srun -c 4 featureCounts -T 4 ... #run mapping and counting in a few lines, doesn't matter the number of samples. for sample in WT1 KO1 ... #deconnect your session logout