Question: Reporting alleles sequences for Amplicon sequencing after variant calling
0
gravatar for Aurelie MLB
4.6 years ago by
Aurelie MLB320
United Kingdom
Aurelie MLB320 wrote:

Hello,

I have amplicon sequencing data for clones whose genome has been edited. On each alleles, there might be one or two indels. I would like report the sequences of the alleles and their frequency. I have some complexe cases where some clones are mixed clones. May I ask if there is an obvious tool to work out what are the amplicon sequences from a variant caller result or do I have to look at all the reads and work out which variants are combined in one sequence?

 

sequencing dna-seq sequence • 1.3k views
ADD COMMENTlink written 4.6 years ago by Aurelie MLB320

@Aurelie MLB I might have very similar problem if not the same. I'm also seeking solution for that. We can kinder of help each other by sharing info. Although this isn't a discussion forum as such, but if we can provide a solution or at least point people in the right direction I think It will super useful to other biologist and bioinformaticians. 

Have a look at this tool https://www.broadinstitute.org/gatk/gatkdocs/org_broadinstitute_gatk_tools_walkers_haplotypecaller_HaplotypeCaller.php 

Also you might need to clarify/explain you question a bit better to me because I'm not exactly sure what you are trying to do...

Here is what I have got: a mouse that had short sequence mutation introduces (~ 26 bp I think (i didn't do the experiment)) Mouse had been sequenced, targeting sequence to the region of the mutation. I can see InDels and other SNP's in IGV. I need reliable method to confidently call and InDel. The problem is not all reads will have a mutation..

I tried that method about and also samtools mpileup and bcftools. I'm still looking for the solution

cheers,

ADD REPLYlink written 4.4 years ago by Kirill270

Hi,

What you want is a variant caller I think. You could check gatk, freebayes, gatk-haplotype, platypus, mutect, scalpel, vardict, varscan, samtools... If you want a tool specifically designed for indels, you could have a look at scalpel, pindel, sid...

My question was a bit different I think as I wanted to find the existing combinations of variants for a short region (to identify the sequence of all alleles). This a step further after variant calling. I did not find an already existing tool doing this really unfortunately.

ADD REPLYlink modified 9 days ago by RamRS24k • written 4.4 years ago by Aurelie MLB320
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