Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • nupurgupta
    Member
    • Aug 2010
    • 29

    vcfutils to get consensus sequence

    There have been several posts about this, but it has been a while, and my queries are not completely answered, so thought I would put it down again.

    I am trying to generate a consensus sequence from a vcf file and a reference genome. I am trying to use vcfutils, and I get an error as

    Use of uninitialized value in addition (+) at /my/bin/vcfutils.pl line 518
    ....

    I am trying to rewrite vcfutils.pl to avoid this error. The error is in the area -

    $q = $1 if ($t[7] =~ /FQ=(-?[\d\.]+)/);
    if ($q < 0) {
    $_ = ($t[7] =~ /AF1=([\d\.]+)/)? $1 : 0;
    $b = ($_ < .5 || $alt eq '.')? $ref : $alt;
    $q = -$q;
    } else {
    $b = $het{"$ref$alt"};
    $b ||= 'N';
    }
    $b = lc($b);
    $b = uc($b) if (($t[7] =~ /MQ=(\d+)/ && $1 >= $_Q) && ($t[7] =~ /DP=(\d+)/ && $1 >= $_d && $1 <= $_D));
    $q = int($q + 33 + .499);
    $q = chr($q <= 126? $q : 126);

    The current format of VCF files does not have an FQ flag, so this code fails when it is trying to parse out the FQ value. My question is - how can I calculate $q from the VCF file?

    There is great information in
    Discussion of next-gen sequencing related bioinformatics: resources, algorithms, open source efforts, etc


    which says
    "$q, which is derived from the FQ, ends up being the quality score"

    But I am still trying to figure out how to calculate this value.

    Thanks very much.
  • Balat
    Member
    • May 2010
    • 36

    #2
    I encountered the same problem with vcfutils.pl

    I ended up using the old (deprecated) pileup file to get the consensus sequence.

    samtools pileup -cf ref.fa aln.bam | samtools.pl pileup2fq -D100 > cns.fastq

    Comment

    • nupurgupta
      Member
      • Aug 2010
      • 29

      #3
      Oh dear..I think the current version of samtools doesnt even support it. Do you know which version you were using, I will get that
      Thanks very much!

      Comment

      • Balat
        Member
        • May 2010
        • 36

        #4
        samtools 1.11

        Comment

        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.
          ...
          Yesterday, 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, 10:04 AM
        0 responses
        11 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 07-08-2026, 10:08 AM
        0 responses
        9 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 07-07-2026, 11:05 AM
        0 responses
        16 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 07-02-2026, 11:08 AM
        0 responses
        31 views
        0 reactions
        Last Post SEQadmin2  
        Working...