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Does anyone have a good handle on the best algorithms to use for mapping/snp calling in very deep coverage of multiple (10 or so) haplotypes in a pooled sample? I'm aware of the MAQ approach and the BWA/samtools approach, but I was surprised by the lack of overlap between these techniques. I also think that they may be confounded by my extremely deep coverage from sequence capture (>200x at most sites). Some of the calls looked a little fishy to me when I pulled them up in IGV, and I'm wondering if something better has come along.

Is there a consensus on the right way to do this, or at least a right way to do this?

(This may be answered in the forums somewhere, but I can't resist trying out the new Q&A format)

asked 21 May '10, 09:22

wjeck's gravatar image

wjeck
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Wow, I wish I had noticed this sooner, but using IGV I now see that my samples are highly contaminated... so much for that data set...

(11 Jun '10, 14:50) wjeck

No, maq/samtools are not designed for deep coverage. I usually recommend people to write their own caller based on the pileup output.

link

answered 21 May '10, 21:13

lh3's gravatar image

lh3
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accept rate: 10%

@lh3 can you provide a link or reference that samtools is not designed for deep coverage? what about varscan?

(16 Jun '10, 23:38) brentp
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@brentp re: lh3. He is the author of samtools ;)

(17 Jun '10, 04:17) nilshomer

@nilshomer , good to know. thanks. :)

(17 Jun '10, 10:13) brentp

The CRISP algorithm by Vikas Bansal for pooled samples might be a way to look at your pooled data. It seems to be designed for higher coverage for the pool, coming down to 20-50x coverage per haplotype

http://bioinformatics.oxfordjournals.org/cgi/content/full/26/12/i318?etoc

"The average coverage of the two pools, based on the alignments, was ~2080x (42x per haplotype) and 2500x (50x per haplotype), respectively "

link

answered 08 Jun '10, 17:40

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bioinfosm
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edited 08 Jun '10, 17:41

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Asked: 21 May '10, 09:22

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Last updated: 02 Feb '11, 15:51

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