#!/bin/bash #This script regenerates test data from replicate RID Q-Y5F6 module load singularity/3.5.3 module load pigz/2.4 ln -sfn /project/BICF/BICF_Core/shared/gudmap/test_data/* ../test_data/ mkdir -p NEW_test_data ln -sfn ./test_data/auth/credential.json ~/.deriva/credential.json mkdir -p ./NEW_test_data/bag singularity run 'docker://gudmaprbk/deriva1.3:1.0.0' deriva-download-cli staging.gudmap.org --catalog 2 ../workflow/conf/Replicate_For_Input_Bag.json . rid=Q-Y5F6 cp Q-Y5F6_inputBag.zip ./NEW_test_data/bag/Q-Y5F6_inputBag_xxxxxxxx.zip mkdir -p ./NEW_test_data/fastq unzip ./NEW_test_data/bag/Q-Y5F6_inputBag_xxxxxxxx.zip singularity run 'docker://gudmaprbk/deriva1.3:1.0.0' bash ../workflow/scripts/bdbag_fetch.sh Q-Y5F6_inputBag Q-Y5F6 cp Q-Y5F6.R1.fastq.gz ./NEW_test_data/fastq/Q-Y5F6.R1.fastq.gz cp Q-Y5F6.R2.fastq.gz ./NEW_test_data/fastq/Q-Y5F6.R2.fastq.gz mkdir -p ./NEW_test_data/fastq/small singularity exec 'docker://gudmaprbk/seqtk1.3:1.0.0' seqtk sample -s100 ./NEW_test_data/fastq/Q-Y5F6.R1.fastq.gz 1000000 1> Q-Y5F6_1M.R1.fastq singularity exec 'docker://gudmaprbk/seqtk1.3:1.0.0' seqtk sample -s100 ./NEW_test_data/fastq/Q-Y5F6.R2.fastq.gz 1000000 1> Q-Y5F6_1M.R2.fastq pigz Q-Y5F6_1M.R1.fastq pigz Q-Y5F6_1M.R2.fastq cp Q-Y5F6_1M.R1.fastq.gz ./NEW_test_data/fastq/small/Q-Y5F6_1M.R1.fastq.gz cp Q-Y5F6_1M.R2.fastq.gz ./NEW_test_data/fastq/small/Q-Y5F6_1M.R2.fastq.gz mkdir -p ./NEW_test_data/meta singularity run 'docker://gudmaprbk/trimgalore0.6.5:1.0.0' trim_galore --gzip -q 25 --illumina --length 35 --basename Q-Y5F6_1M.se -j 20 ./NEW_test_data/fastq/small/Q-Y5F6_1M.R1.fastq.gz singularity run 'docker://gudmaprbk/trimgalore0.6.5:1.0.0' trim_galore --gzip -q 25 --illumina --length 35 --paired --basename Q-Y5F6_1M.pe -j 20 ./NEW_test_data/fastq/small/Q-Y5F6_1M.R1.fastq.gz ./NEW_test_data/fastq/small/Q-Y5F6_1M.R2.fastq.gz cp Q-Y5F6_1M.se_trimmed.fq.gz ./NEW_test_data/fastq/small/Q-Y5F6_1M.se_trimmed.fq.gz cp Q-Y5F6_1M.pe_val_1.fq.gz ./NEW_test_data/fastq/small/Q-Y5F6_1M.pe_val_1.fq.gz cp Q-Y5F6_1M.pe_val_2.fq.gz ./NEW_test_data/fastq/small/Q-Y5F6_1M.pe_val_2.fq.gz cp Q-Y5F6_1M.R1.fastq.gz_trimming_report.txt ./NEW_test_data/meta/Q-Y5F6_1M.R1.fastq.gz_trimming_report.txt cp Q-Y5F6_1M.R2.fastq.gz_trimming_report.txt ./NEW_test_data/meta/Q-Y5F6_1M.R2.fastq.gz_trimming_report.txt touch metaTest.csv echo 'Replicate_RID,Experiment_RID,Study_RID,Paired_End,File_Type,Has_Strand_Specific_Information,Used_Spike_Ins,Species,Read_Length' > metaTest.csv echo 'Replicate_RID,Experiment_RID,Study_RID,uk,FastQ,no,no,Homo sapiens,75' >> metaTest.csv cp metaTest.csv ./NEW_test_data/meta/metaTest.csv mkdir -p ./NEW_test_data/bam mkdir -p ./NEW_test_data/bam/small singularity run 'docker://gudmaprbk/hisat2.2.1:1.0.0' hisat2 -p 20 --add-chrname --un-gz Q-Y5F6_1M.se.unal.gz -S Q-Y5F6_1M.se.sam -x /project/BICF/BICF_Core/shared/gudmap/references/new/GRCm38.p6.vM25/hisat2/genome --rna-strandness F -U ./NEW_test_data/fastq/small/Q-Y5F6_1M.se_trimmed.fq.gz --summary-file Q-Y5F6_1M.se.alignSummary.txt --new-summary singularity run 'docker://gudmaprbk/hisat2.2.1:1.0.0' samtools view -1 -@ 20 -F 4 -F 8 -F 256 -o Q-Y5F6_1M.se.bam Q-Y5F6_1M.se.sam singularity run 'docker://gudmaprbk/hisat2.2.1:1.0.0' samtools sort -@ 20 -O BAM -o Q-Y5F6_1M.se.sorted.bam Q-Y5F6_1M.se.bam singularity run 'docker://gudmaprbk/hisat2.2.1:1.0.0' samtools index -@ 20 -b Q-Y5F6_1M.se.sorted.bam Q-Y5F6_1M.se.sorted.bam.bai singularity run 'docker://gudmaprbk/hisat2.2.1:1.0.0' hisat2 -p 20 --add-chrname --un-gz Q-Y5F6_1M.pe.unal.gz -S Q-Y5F6_1M.pe.sam -x /project/BICF/BICF_Core/shared/gudmap/references/new/GRCm38.p6.vM25/hisat2/genome --rna-strandness FR --no-mixed --no-discordant -1 ./NEW_test_data/fastq/small/Q-Y5F6_1M.pe_val_1.fq.gz -2 ./NEW_test_data/fastq/small/Q-Y5F6_1M.pe_val_2.fq.gz --summary-file Q-Y5F6_1M.pe.alignSummary.txt --new-summary singularity run 'docker://gudmaprbk/hisat2.2.1:1.0.0' samtools view -1 -@ 20 -F 4 -F 8 -F 256 -o Q-Y5F6_1M.pe.bam Q-Y5F6_1M.pe.sam singularity run 'docker://gudmaprbk/hisat2.2.1:1.0.0' samtools sort -@ 20 -O BAM -o Q-Y5F6_1M.pe.sorted.bam Q-Y5F6_1M.pe.bam singularity run 'docker://gudmaprbk/hisat2.2.1:1.0.0' samtools index -@ 20 -b Q-Y5F6_1M.pe.sorted.bam Q-Y5F6_1M.pe.sorted.bam.bai cp Q-Y5F6_1M.se.bam ./NEW_test_data/bam/small/Q-Y5F6_1M.se.bam cp Q-Y5F6_1M.pe.bam ./NEW_test_data/bam/small/Q-Y5F6_1M.pe.bam cp Q-Y5F6_1M.se.sorted.bam ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.bam cp Q-Y5F6_1M.se.sorted.bam.bai ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.bam.bai cp Q-Y5F6_1M.pe.sorted.bam ./NEW_test_data/bam/small/Q-Y5F6_1M.pe.sorted.bam cp Q-Y5F6_1M.pe.sorted.bam.bai ./NEW_test_data/bam/small/Q-Y5F6_1M.pe.sorted.bam.bai cp Q-Y5F6_1M.se.alignSummary.txt ./NEW_test_data/meta/Q-Y5F6_1M.se.alignSummary.txt cp Q-Y5F6_1M.pe.alignSummary.txt ./NEW_test_data/meta/Q-Y5F6_1M.pe.alignSummary.txt singularity run 'docker://gudmaprbk/picard2.23.9:1.0.0' java -jar /picard/build/libs/picard.jar MarkDuplicates I=./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.bam O=Q-Y5F6_1M.se.deduped.bam M=Q-Y5F6_1M.se.deduped.Metrics.txt REMOVE_DUPLICATES=true singularity run 'docker://gudmaprbk/picard2.23.9:1.0.0' samtools sort -@ 20 -O BAM -o Q-Y5F6_1M.se.sorted.deduped.bam Q-Y5F6_1M.se.deduped.bam singularity run 'docker://gudmaprbk/picard2.23.9:1.0.0' samtools index -@ 20 -b Q-Y5F6_1M.se.sorted.deduped.bam Q-Y5F6_1M.se.sorted.deduped.bam.bai cp Q-Y5F6_1M.se.deduped.bam ./NEW_test_data/bam/small/Q-Y5F6_1M.se.deduped.bam cp Q-Y5F6_1M.se.sorted.deduped.bam ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.bam cp Q-Y5F6_1M.se.sorted.deduped.bam.bai ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.bam.bai cp Q-Y5F6_1M.se.deduped.Metrics.txt ./NEW_test_data/meta/Q-Y5F6_1M.se.deduped.Metrics.txt for i in {"chr8","chr4","chrY"}; do echo "samtools view -b ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.bam ${i} > Q-Y5F6_1M.se.sorted.deduped.${i}.bam; samtools index -@ 20 -b Q-Y5F6_1M.se.sorted.deduped.${i}.bam Q-Y5F6_1M.se.sorted.deduped.${i}.bam.bai;"; done | singularity run 'docker://gudmaprbk/picard2.23.9:1.0.0' parallel -j 20 -k cp Q-Y5F6_1M.se.sorted.deduped.chr4.bam ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.chr4.bam cp Q-Y5F6_1M.se.sorted.deduped.chr4.bam.bai ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.chr4.bam.bai cp Q-Y5F6_1M.se.sorted.deduped.chr8.bam ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.chr8.bam cp Q-Y5F6_1M.se.sorted.deduped.chr8.bam.bai ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.chr8.bam.bai cp Q-Y5F6_1M.se.sorted.deduped.chrY.bam ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.chrY.bam cp Q-Y5F6_1M.se.sorted.deduped.chrY.bam.bai ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.chrY.bam.bai mkdir -p ./NEW_test_data/counts mkdir -p ./NEW_test_data/counts/small ln -s /project/BICF/BICF_Core/shared/gudmap/references/new/GRCm38.p6.vM25/metadata/geneID.tsv ln -s /project/BICF/BICF_Core/shared/gudmap/references/new/GRCm38.p6.vM25/metadata/Entrez.tsv singularity run 'docker://gudmaprbk/subread2.0.1:1.0.0' featureCounts -T 20 -a /project/BICF/BICF_Core/shared/gudmap/references/new/GRCm38.p6.vM25/annotation/genome.gtf -G /project/BICF/BICF_Core/shared/gudmap/references/new/GRCm38.p6.vM25/sequence/genome.fna -g 'gene_name' --extraAttributes 'gene_id' -o Q-Y5F6_1M.se_countData -s 1 -R SAM --primary --ignoreDup ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.bam singularity run 'docker://gudmaprbk/subread2.0.1:1.0.0' Rscript ../workflow/scripts/calculateTPM.R --count Q-Y5F6_1M.se_countData singularity run 'docker://gudmaprbk/subread2.0.1:1.0.0' Rscript ../workflow/scripts/convertGeneSymbols.R --repRID Q-Y5F6_1M.se cp Q-Y5F6_1M.se_countData ./NEW_test_data/counts/small/Q-Y5F6_1M.se_countData cp Q-Y5F6_1M.se.countTable.csv ./NEW_test_data/counts/small/Q-Y5F6_1M.se.countTable.csv cp Q-Y5F6_1M.se_tpmTable.csv ./NEW_test_data/counts/small/Q-Y5F6_1M.se_tpmTable.csv mkdir -p ./NEW_test_data/bw mkdir -p ./NEW_test_data/bw/small singularity run 'docker://gudmaprbk/deeptools3.5.0:1.0.0' bamCoverage -p 20 -b ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.bam -o Q-Y5F6_1M.se.bw cp Q-Y5F6_1M.se.bw ./NEW_test_data/bw/small/Q-Y5F6_1M.se.bw mkdir -p ./NEW_test_data/fastqc mkdir -p ./NEW_test_data/fastqc/small singularity run 'docker://gudmaprbk/fastqc0.11.9:1.0.0' fastqc ./NEW_test_data/fastq/small/Q-Y5F6_1M.R1.fastq.gz -o . cp Q-Y5F6_1M.R1_fastqc.html ./NEW_test_data/fastqc/small/Q-Y5F6_1M.R1_fastqc.html cp Q-Y5F6_1M.R1_fastqc.zip ./NEW_test_data/fastqc/small/Q-Y5F6_1M.R1_fastqc.zip echo -e "geneID\tchrom\ttx_start\ttx_end\tTIN" > Q-Y5F6_1M.se.sorted.deduped.tin.xls for i in {"chr8","chr4","chrY"}; do echo "tin.py -i ./NEW_test_data/bam/small/Q-Y5F6_1M.se.sorted.deduped.${i}.bam -r /project/BICF/BICF_Core/shared/gudmap/references/new/GRCm38.p6.vM25/annotation/genome.bed; cat Q-Y5F6_1M.se.sorted.deduped.${i}.tin.xls | tr -s \"\\w\" \"\\t\" | grep -P \"\\t${i}\\t\";"; done | singularity run 'docker://gudmaprbk/rseqc4.0.0:1.0.0' parallel -j 20 -k >> Q-Y5F6_1M.se.sorted.deduped.tin.xls cp Q-Y5F6_1M.se.sorted.deduped.tin.xls ./NEW_test_data/meta/Q-Y5F6_1M.se.sorted.deduped.tin.xls chgrp -R BICF_Core ./NEW_test_data chmod -R 750 ./NEW_test_data