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Genotype SNP with IGV

Hi

I have bam files of WXS and I'm visualizing it with IGV. Can I genotype a SNP only by visualizing its reads and bases in IGV? Are there any standard? Like minimum coverage or a threshould to call heterozygous (20%? 30%?).

Example: In some snp, in igv, I have a 20x coverage for that position and 20% of C and 80% of T (observing homogeneous distribution in both strands) . Can I call this a CT?

Are there any other way to call a genotype of a SNP in NGS data (bam) other than variant caller? I did that, but some of my snps are not being called in VCF file.

Thanks.

igv ngs snp

1 answer

There is no standard for this, and performing this manual inspection requires experience more than anything else. However, as I recently did this for a private company in New York, USA, I can share my own experience.

That of which to be aware:

  • overall read depth over the variant position. For a germline variant, technically, my work from the clinical setting in the NHS England shows that 18 is the bare minimum that one should want, with 30 being ideal.
  • read bias - is the variant mostly called in R1 or R2?
  • position bias - is the variant called at the end of reads?
  • quality of reads - are the reads flagged in IGV for any reason?
  • MAPQ of the reads
  • are there many base-mismatches in the reads aligning to the general region surrounding the variant, and/or indels with a seemingly sporadic / random distribution?
  • is it a repeat region, or do any reads span a repeat region by even a few bases?

Probably more criteria about which to be aware.

Kevin

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