Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • samtools mpileup filter SNPs

    I used samtools mpileup to call SNPs and indels, followed by vcfutils.pl, as suggested in http://samtools.sourceforge.net/mpileup.shtml

    I wonder if there are additional filters that I should use for exome data. I saw in publication people use for example, ">=3 reads support a variation", ">=5% on both strands", "consensus quality >=20". I know "-d3" can be used to assign a minimum number of reads, but can anyone tell me how to set filters for the strand bias? And is the 6th column (QUAL) the "consensus quality" which should be required to be >=20?

    Any help is appreciated! Thank you

  • #2
    Maybe vcfutils.pl varFilter?

    Comment


    • #3
      Yes I used varFilter -- I forgot to mention this. I followed the mpileup webpage and used the following two lines:

      samtools mpileup -uf ref.fa aln1.bam aln2.bam | bcftools view -bvcg - > var.raw.bcf
      bcftools view var.raw.bcf | vcfutils.pl varFilter -D10000 > var.flt.vcf

      I have exome data so I used -D10000, but I'm not sure if there are other rules that should be used, especially for those decribed in publications like strand bias (mentioned above).

      Comment


      • #4
        I was thinking specifically of this option

        -1 FLOAT min P-value for strand bias (given PV4) [0.0001]

        Strand bias is not something I've thought much (at all) about, but this option seems like it might be worth following up.

        Comment

        Latest Articles

        Collapse

        • seqadmin
          Current Approaches to Protein Sequencing
          by seqadmin


          Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
          04-04-2024, 04:25 PM
        • seqadmin
          Strategies for Sequencing Challenging Samples
          by seqadmin


          Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
          03-22-2024, 06:39 AM

        ad_right_rmr

        Collapse

        News

        Collapse

        Topics Statistics Last Post
        Started by seqadmin, 04-11-2024, 12:08 PM
        0 responses
        22 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 04-10-2024, 10:19 PM
        0 responses
        24 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 04-10-2024, 09:21 AM
        0 responses
        20 views
        0 likes
        Last Post seqadmin  
        Started by seqadmin, 04-04-2024, 09:00 AM
        0 responses
        52 views
        0 likes
        Last Post seqadmin  
        Working...
        X