SEQanswers

SEQanswers (http://seqanswers.com/forums/index.php)
-   Bioinformatics (http://seqanswers.com/forums/forumdisplay.php?f=18)
-   -   generating pileup of paired end fragments (http://seqanswers.com/forums/showthread.php?t=49678)

Slacanch 01-20-2015 07:39 AM

generating pileup of paired end fragments
 
Dear all,
i'm working with paired end rnaseq data. i wanted to generate a pileup with samtools mpileup but noticed that only the mapping reads get counted, and not the unmapped part between two mates, like so:

______READ1--------READ2________ <- mapping
______1 1 1 1 0 0 0 1 1 1 1________ <- pileup

is generating pileups using the whole fragment (read + mate + part in between them) frowned upon for some reason? would it be correct to do it? if yes are there any tools?

alternatively, is there a tool to merge mates and generate the complete fragment in a sam file?

Thank you!

dpryan 01-20-2015 07:55 AM

What would a pileup of the uncovered region between mates even look like. They couldn't contribute any sequence, so what would you show for them? The only conceivable reason I could think of to do this is in peak calling for chip-seq or something like that, but then you don't care about the actual sequence so there'd be no point in going to full pileup route.

Slacanch 01-20-2015 08:20 AM

well i don't really care about the sequence, it's for profile comparison.
as long as the positions are right, i'm ok with it.

maybe i shouldn't have called it pileup, but coverage. just a measure of how many reads overlap each position, in order to be able to compare profiles at certain loci (such as transcriptional readthrough for example).

for future reference, i discovered a script that does more or less this in the "misc" folder of samtools. it's called interpolate_sam.pl.

i'll check it more in detail and report back.


All times are GMT -8. The time now is 09:58 AM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2019, vBulletin Solutions, Inc.