Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • Mark
    Member
    • Nov 2008
    • 54

    PacBio software vcf and consensus fasta do not agree

    I'm using PacBio's "variantCaller" tool (v6) to call variants on some older RSII subread data I aligned with blasr. The reference is 14845 bp.

    My variantCaller command is:

    Code:
    variantCaller --algorithm arrow --log-file variantCaller.log --annotateGFF --reportEffectiveCoverage --noEvidenceConsensusCall lowercasereference --minCoverage 40 --coverage 100 --minMapQV 40 --minConfidence 10 --minReadScore 0.75 --minSnr 3.75 --minZScore -3.5 --minAccuracy 0.82  --numWorkers 5 -r ref.fa -o 129C02-vs-ref.fa.sort.vcf -o 129C02-vs-ref.fa.sort.gff -o 129C02-vs-ref.fa.sort.fasta -o 129C02-vs-ref.fa.sort.fastq 129C02-vs-ref.fa.sort.bam
    Now it is my understanding that the fasta (and fastq) output this command should reflect the differences between the reference and the reads as reported in the vcf output.

    My vcf file reports:

    Code:
    #CHROM  POS     ID      REF     ALT     QUAL    FILTER  INFO
    ref    11438   .       CA      C       25      PASS    DP=100
    However when I align the reference and this fasta output 2 1-bp deletions are observed. One is the deletion in the vcf. The other is several thousand base pairs away and is not called by variantCaller.

    This, of course, is very disturbing. Can you explain this?

    Thanks for your help

Latest Articles

Collapse

  • SEQadmin2
    Advanced Sequencing Platforms Tackle Neuroscience’s Toughest Genomics Problems
    by SEQadmin2



    Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
    ...
    07-09-2026, 11:10 AM
  • SEQadmin2
    Cancer Drug Resistance: The Lingering Barrier to Rising Survival
    by SEQadmin2



    Cancer survival rates have significantly increased in the last few decades in the United States, reaching a combined 70% 5-year survival rate by 2021. Behind this number, there are years of research to find new therapies, drug targets, and early detection methods. But there is one core challenge that keeps slowing down these advances, and it’s about drug resistance.

    There is no single reason why many patients don’t respond to treatment as expected. Cancer is...
    07-08-2026, 05:17 AM
  • GATTACAT
    Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
    by GATTACAT
    Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
    07-01-2026, 11:43 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by SEQadmin2, Yesterday, 10:26 AM
0 responses
12 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 07-09-2026, 10:04 AM
0 responses
25 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 07-08-2026, 10:08 AM
0 responses
16 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 07-07-2026, 11:05 AM
0 responses
33 views
0 reactions
Last Post SEQadmin2  
Working...