Question: Variant Allele Frequency from Ion Torrent data
3
gravatar for stamakro
5.4 years ago by
stamakro40
Netherlands
stamakro40 wrote:

I have a vcf file with SNPs called using the Torrent Variant Caller and I want to calculate their Variant Allele Frequencies. 

The issue is that there is both a DP (depth) and a FPD (flow depth) fields as well as an AO (Alternate Allele Observations) and a FAO (Flow Alternate Allele Observations) fields and the two often disagree by a lot.

Which pair should I trust? DP & AO or FDP & FAO?

Or should I just ignore the cases where they disagree?

Thanks
 

sequencing snp vaf ion torrent • 3.1k views
ADD COMMENTlink modified 3.6 years ago by Biostar ♦♦ 20 • written 5.4 years ago by stamakro40
6
gravatar for Katie D'Aco
5.4 years ago by
Katie D'Aco1.0k
Massachusetts
Katie D'Aco1.0k wrote:

The fields FPD and FAO are after correcting for flow order of nucleotides, and so should be more accurate.

From https://ionreporter.thermofisher.com/ionreporter/help/GUID-A6D3AE1A-84C6-4B66-AA0D-69244576D2FE.html it looks like they don't use FPD in their allele freq calculations:

"AF=FAO/(FAO+FRO) and not FAO/FDP. This is because FDP may include reads that don't fit the flow space profile of any hypothesis; in such cases, FDP>=FAO+FRO and this is not used in allele frequency calculation"

ADD COMMENTlink written 5.4 years ago by Katie D'Aco1.0k
Please log in to add an answer.

Help
Access

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.
Powered by Biostar version 2.3.0
Traffic: 2440 users visited in the last hour
_