Publication: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6707278/ PRJNA517683 #!/bin/bash #SBATCH --time=23:00:00 #SBATCH --nodes=1 #SBATCH --ntasks-per-node=36 #SBATCH --job-name="get1" #SBATCH --mail-type=BEGIN #SBATCH --mail-type=END #SBATCH --mail-type=FAIL #SBATCH --output=job.%J.out #SBATCH --error=job.%J.err #SBATCH --mail-user=mhrwoodhouse@gmail.com #SBATCH --mail-type=BEGIN,END,FAIL ulimit -s unlimited module load trimgalore module load bowtie module load samtools module load picard module load bedtools2 for sample in *.fastq.gz do echo $sample describer=$(echo ${sample} | sed 's/.fastq.gz//') echo $describer trim_galore ${sample} zcat ${describer}_trimmed.fq.gz > ${describer}_trimmed.fq bowtie -X 1000 -m 1 -v 2 --best --strata /work/LAS/mhufford-lab/mw2/epigenetics/bowtie_B73v5/B73_v5 ${describer}_trimmed.fq -S ${describer}_trimmed.sam samtools view --threads 36 -b -o ${describer}_trimmed.bam ${describer}_trimmed.sam samtools sort -o ${describer}_sorted.bam -T ${sample}_temp --threads 36 ${describer}_trimmed.bam picard MarkDuplicates I=${describer}_sorted.bam O=${describer}_removed_dups.bam M=marked_dup_metrics.txt REMOVE_DUPLICATES=true bedtools genomecov -bg -ibam ${describer}_removed_dups.bam -split -scale 1.0 > ${describer}.bedgraph /work/LAS/mhufford-lab/mw2/kentUtils/bin/linux.x86_64/bedGraphToBigWig ${describer}.bedgraph /work/LAS/mhufford-lab/mw2/epigenetics/B73v5.sizes ${describer}.bw done For merging the tassel2 and 3 reads: #!/bin/bash #SBATCH --time=05:00:00 #SBATCH --nodes=1 #SBATCH --ntasks-per-node=36 #SBATCH --job-name="merge" #SBATCH --mail-type=BEGIN #SBATCH --mail-type=END #SBATCH --mail-type=FAIL #SBATCH --output=job.%J.out #SBATCH --error=job.%J.err #SBATCH --mail-user=mhrwoodhouse@gmail.com #SBATCH --mail-type=BEGIN,END,FAIL module load samtools module load bedtools2 module load picard For calling peaks with peakranger: /work/LAS/mhufford-lab/mw2/epigenetics/peakranger ranger --format bam SRR8502759_removed_dups.bam SRR8502757_removed_dups.bam SRR8502759_rep2_ranger -t 15 /work/LAS/mhufford-lab/mw2/epigenetics/peakranger ranger --format bam SRR8502760_removed_dups.bam SRR8502758_removed_dups.bam SRR8502760_rep2_ranger -t 15 for calling peaks with MACS2: macs2 callpeak -t SRR8502759_removed_dups.bam -c SRR8502757_removed_dups.bam -f BAMPE -n SRR8502759_rep2_macs2 -g 2.2e9 --keep-dup all macs2 callpeak -t SRR8502760_removed_dups.bam -c SRR8502758_removed_dups.bam -f BAMPE -n SRR8502760_rep1_macs2 -g 2.2e9 --keep-dup all