Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • ErikFas
    Member
    • Jun 2014
    • 86

    #1

    Variant calling with GATK between two cell lines

    I recently got some whole genome sequencing data from my platform that would constitute some validation data for my other RNA-seq data. The idea was that the platform would do all the bioinformatic analyses for this data, so that I could keep my focus on RNA rather than DNA, but it turns out that they don't routinely do the last part of the analysis that I want: finding any differing mutations between my two sample types. I now have to do that myself, so I come here for help.

    I have two isogenic cell lines, theoretically differing in a single mutation, and that is what we want to confirm (or that, at least, any other mutatations are in non-functional or otherwise non-relevant regions). As far as I understand it, the platform has done the alignment and the heaviest parts of the analysis, as well as a variant calling relative the reference. I'm really only interested in the difference between the cell lines, and not between cell lines / reference.

    I was pointed towards the GenotypeGVCFs function of GATK. If I understand it correctly, what I do is input my two samples GVCFs and run it. I'm not sure why I need a reference, though. I made some rudimentary code:

    Code:
    java -jar GenomeAnalysisTK.jar \
    	-T GenotypeGVCFs \
            -nt 16 \
    	-R .../human_g1k_v37.fasta \
    	--variant .../sample1.clean.dedup.recal.bam.genomic.vcf \
    	--variant .../sample2.clean.dedup.recal.bam.genomic.vcf \
    	-o .../output.vcf
    Is this what I'm looking for? If not, what am I missing, or am I completely off course?

Latest Articles

Collapse

  • kinpict.com
    Reply to 16S/ITS Profiling vs. Shotgun Metagenomics: What Changes in Your Workflow?
    by kinpict.com
    The part about this topic feels surprisingly grounded, and the details on the page is the bit that made me slow down. I’d keep <a href="https://kinpict.com/">ai family photo generator</a> bookmarked for later.
    Today, 06:37 PM
  • SEQadmin2
    16S/ITS Profiling vs. Shotgun Metagenomics: What Changes in Your Workflow?
    by SEQadmin2





    NGS has revolutionized clinical microbiology by offering new ways to identify and surveil pathogens without prior knowledge of the target. One popular approach is 16S rRNA and ITS amplicon sequencing, which targets specific microbial markers. But studies are increasingly incorporating shotgun metagenomics, a technique that sequences all DNA from a sample without targeting specific microbial markers. This approach allows more detailed taxonomic classification,...
    10-30-2026, 06:46 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by SEQadmin2, 11-12-2026, 07:59 AM
0 responses
27 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 11-05-2026, 09:25 AM
0 responses
35 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 11-03-2026, 11:54 AM
2 responses
56 views
0 reactions
Last Post kinpict.com  
Started by SEQadmin2, 10-28-2026, 09:57 AM
0 responses
175 views
0 reactions
Last Post SEQadmin2  
Working...