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Showing content from https://github.com/samtools/samtools/issues/2210 below:

samtools stats - Dovetailed read pairs orientation · Issue #2210 · samtools/samtools · GitHub

I found that samtools stats classifies dovetailed reads (share the same leftmost position) are classified as either outward- or inward-oriented depending on if read1 (blue in the image below) is on the forward or reverse strand.

The relevant code can be found here: https://github.com/samtools/samtools/blob/4224ee098020b8957bf19343ecbd45f9d7e76efa/stats.c#L1257C13-L1263C14

This results in that dovetailed read pairs are evenly split between being classified as outward or inward.

% samtools view -bh P33410_1001.bam -e 'pos == mpos' | samtools stats | grep orient 
SN	inward-oriented pairs:	245641
SN	outward oriented pairs:	243434 

For me, it would make more sense that these reads should rather all be classified as inward-oriented as the ends in fact do not protrude beyond the mate. What do you think?


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