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MACS peak calling of paired-end reads in SAM format

HI,

Just a quick question: if SAM format is used for paired-end reads, will these be automatically recognised as paired-end by MACS when doing peak-calling?

Thanks!

chip-seq macs sam

I am confused since MACS2 run with default parameters writes in the report file "# Paired-End mode is off"

I am running it as follows: macs2 callpeak -t chip.sam -c input.sam --broad -g hs --broad-cutoff 0.1

1 answer

MACS was developed in the days of short single-end reads. As far as I know, it can autodetect the file type (SAM, BAM, BED, Eland...), but you have to indicate if paired-end reads are present of not. I would write your SAM files as BAM (samtools view -b in.sam -o out.bam), and then add to your command -f BAMPE. You can also check from the stuff MACS writes to STDERR if MACS says "read treatment tags" (indicating single end) or "read treatment fragments" (indicating paired-reads, because two reads are read that are sequenced from one fragment).

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