Samtools Snp Calling Problem
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12.9 years ago
Liyf ▴ 300

Hi, everyone. I found a strange thing when doing snp calling using my exome sequencing data. I use BWA to map my reads to human genome hg19 and samtools whith default parameters to call snps. But when completed, I use samtools mpileup command, and accidently found that some positions, which are obvious a heterozygosis, but not in my SNP lists. I do not know why.

chr7    128141865   N   412 T$cTTTCCCCTtCTcCTCTTCCcTTTCCCCTCTtCCTTCTCcTTcCCTTTCCTTTTTTTTCCTTTCtcCCCTCCTtCTCTTcCCCTCtCTTCCTcTcCTTTcTTtTCTTccCCTCCCCCCTTcTCCCCCCttTtctTTCTCTTTcTTCTCCCTCTTCTCTCTTCcCTTTtcTTCTTTtTCTTCCTCCTTTCTTTccTCCCTTCCTTTCCTTTcctTCTCCccCTCCTCCcttCTCCtCTTTCCTtTTTtCCtcTCCTCTtTTtcCTtCTCcccCCTTCtCTTtcTCTTttCTCCCTcCCCtTCTcTTTcctCTTTTTCTccTCCCtccCTTTttcTCCccCCTCTCTttTCCctCTCCccCCtcttcttcCCCCccTCTctttCCTtTCctCCTCTtcCTcccTCTCCCc^]t^]t^]
>CDDDDDDD@F9DFDADCDCDHDDDCCDDCDDFCCCD?DDGDDECEADE>EEEECECEE>ECCEEGJFDFDFFC2FB@DDADF;EHJHE>EHHGHJHHHG8G>IHHHFGJJJ:EJJGJIJIDJJIG>EJI@J(HCIJJGIJEGCIIJJJJI@JICIIIJJH=CFGB:B2E@JIJIJFJJIGHHIJJGGIHIJ8;FHJHHHIHJHBHI9GG=FFDIJI>BDDJHHJJDFCDEJGC@BJFJJJJG<IIHBJ@BDIJJGIHBJJ?=GBDJJJ?DDJIGJIBJGBDAIGJHAAGJJIJADIJJDIJCBJJIDD<DIJHHEEIDDJFJIBDDJIJJDB7JJJDDJJ2EGJGBBJJJ?(JJHH@<HHD@@BDB?DHHHHD?HHD(BB@FDFBFFD<FFFDDA?BFDBDCC@@CCB8?@
snp snp samtools • 3.6k views
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It might help if you could provide the exact commands you used. Thanks.

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The sequence formatting of your single vcf is not very helpful (as you can easily see it breaks layout completely). Further, you need to provide more evidence, about why you think, which location is the one in question and why it is 'obvious a heterozygous'. I couldn't see that at all.

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Best to format long sequence as 'code', fixed.

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I don't see the output of variant calling, just the mpileup output. Could you add the VCF output, also, as well as the command-line you used to produce it?

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I just use the default parameters.

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Michael, I just see the whole chr7 using mpileup, and using perl to identify heterozygous, this position just one of some heterozygous.I say it is heterozygous, because it has C and T both.

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Frozenwithjoy, thank you for your kind help. I just use the default parameters on the samtools website.

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Sean, the VCF output has no this position. Also, the SNPs I call are almost right when I check using tview command. But just like this one, tview tells me that it should be a SNP, but it is not in my result. . ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, . ,,,c,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, . ...C.............................................. ,,,,,,,,,,,,,,,,,,,,,,,,,,, .. ,,c,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, .. ,,c,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,

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Sean, the VCF output has no this position. Also, the SNPs I call are almost right when I check using tview command. But just like this one, tview tells me that it should be a SNP, but it is not in my result.

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I checked it, and found there are no indels around.

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