#!/bin/bash rawdata=../rawdata/atac-seq metadata=../metadata/atac-seq reference=../metadata/reference/hg19_insert result=../result/atac-seq fastq_merge() { cat ${rawdata}/${1}_L3_R1.fq.gz ${rawdata}/${1}_L4_R1.fq.gz > ${metadata}/${1}_R1.fastq.gz cat ${rawdata}/${1}_L3_R2.fq.gz ${rawdata}/${1}_L4_R2.fq.gz > ${metadata}/${1}_R2.fastq.gz } pre_mapping() { fastp -w 4 -c -h ${metadata}/${1}.fastp.html -j ${metadata}/${1}.fastp.json \ -i ${metadata}/${1}_R1.fastq.gz -o ${metadata}/${1}_r1.fq.gz \ -I ${metadata}/${1}_R2.fastq.gz -O ${metadata}/${1}_r2.fq.gz } mapping() { bowtie2 -p 20 -t --mm --very-sensitive -x ${reference} \ -1 ${metadata}/${1}_r1.fq.gz \ -2 ${metadata}/${1}_r2.fq.gz | samtools view -bS - > ${metadata}/${1}.bam samtools sort -n -o ${metadata}/${1}_nsort.bam ${metadata}/${1}.bam samtools fixmate -m ${metadata}/${1}_nsort.bam ${metadata}/${1}_nsort_fix.bam samtools sort -o ${metadata}/${1}_fix_sort.bam ${metadata}/${1}_nsort_fix.bam samtools index ${metadata}/${1}_fix_sort.bam } post_mapping() { picard MarkDuplicates I=${metadata}/${1}_fix_sort.bam \ O=${metadata}/${1}_sort_mdup.bam M=${metadata}/${1}_mdup_metrics.txt samtools view -F 1804 -f 2 -q 1 -u ${metadata}/${1}_sort_mdup.bam | samtools sort \ -o ${metadata}/${1}_rmdup_sort.bam - samtools index ${metadata}/${1}_rmdup_sort.bam samtools view -h ${metadata}/${1}_rmdup_sort.bam | grep -v -P '\tchrM\t' | samtools view \ -b - > ${metadata}/${1}_sort_rm.bam samtools index ${metadata}/${1}_sort_rm.bam } peak_calling() { macs2 callpeak -t ${metadata}/${1}_sort_rm.bam -n ${1} --outdir ${metadata} -g hs -f BAM --nomodel --shift -100 --extsize 200 -B --SPMR bedGraphToBigWig ${metadata}/${1}_treat_pileup.bdg ../metadata/reference/hg19_insert.chromsize ${metadata}/${1}.bw } quality_report() { ataqv --threads 20 --peak-file ${metadata}/${1}_peaks.narrowPeak human ${metadata}/${1}_sort_mdup.bam --metrics-file ${metadata}/${1}.ataqv.json.gz mkarv ${metadata}/${1} ${metadata}/${1}.ataqv.json.gz } scale_factor() { multiBamSummary bins -b ${metadata}/*_sort_rm.bam --smartLabels -p 20 \ --scalingFactors ${metadata}/scalingFactors.txt -o ${metadata}/readCounts.npz } get_coverage() { bamCoverage -b ${metadata}/high_sort_rm.bam \ -o ${metadata}/high.SeqDepthNorm.bw -e -p 5 --scaleFactor 1.0903 \ --binSize 25 bamCoverage -b ${metadata}/low_sort_rm.bam \ -o ${metadata}/low.SeqDepthNorm.bw -e -p 5 --scaleFactor 1.1836 \ --binSize 25 bamCoverage -b ${metadata}/A1_sort_rm.bam \ -o ${metadata}/A1.SeqDepthNorm.bw -e -p 5 --scaleFactor 0.7303 \ --binSize 25 } for prefix in high low A1; do { fastq_merge "${prefix}" > "${result}/${prefix}.fastq_merge.log" 2>&1 pre_mapping "${prefix}" > "${result}/${prefix}.pre_mapping.log" 2>&1 mapping "${prefix}" > "${result}/${prefix}.mapping.log" 2>&1 post_mapping "${prefix}" > "${result}/${prefix}.post_mapping.log" 2>&1 peak_calling "${prefix}" > "${result}/${prefix}.peak_calling.log" 2>&1 quality_report "${prefix}" > "${result}/${prefix}.quality_report.log" 2>&1 } & done scale_factor > "${result}/scale_factor.log" 2>&1 get_coverage > "${result}/get_coverage.log" 2>&1