Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • SNP at every chromosome position

    Hi,

    I SOLiD sequencing data. After aligning with SHRiMP2, I used samtools mpileup for SNP calling:

    samtools mpileup -C50 -gf hg38.fa -o var.raw.bcf input.bam

    bcftools call -o var.raw.vcf -O v -c var.raw.bcf

    The raw vcf file looks like this:

    #CHROM POS ID REF ALT QUAL FILTER INFO FORMAT SINDHI
    chr1 33953 . T . 56.8087 PASS DP=11;MQSB=0.950952;MQ0F=1;AF1=0;AC1=0;DP4=6,5,0,0;MQ=0;FQ=-59.9998 GT:PL 0/0:0
    chr1 33954 . C . 56.8087 PASS DP=11;MQSB=0.950952;MQ0F=1;AF1=0;AC1=0;DP4=6,5,0,0;MQ=0;FQ=-59.9998 GT:PL 0/0:0
    chr1 33955 . T . 56.4609 PASS DP=10;MQSB=0.952347;MQ0F=1;AF1=0;AC1=0;DP4=5,5,0,0;MQ=0;FQ=-56.997 GT:PL 0/0:0
    chr1 35396 . C . 56.4609 PASS DP=10;MQSB=0.952347;MQ0F=1;AF1=0;AC1=0;DP4=5,5,0,0;MQ=0;FQ=-56.997 GT:PL 0/0:0
    chr1 35397 . C . 61.2368 PASS DP=12;MQSB=0.95494;MQ0F=1;AF1=0;AC1=0;DP4=5,7,0,0;MQ=0;FQ=-62.9905 GT:PL 0/0:0
    chr1 35398 . C . 61.2368 PASS DP=12;MQSB=0.95494;MQ0F=1;AF1=0;AC1=0;DP4=5,7,0,0;MQ=0;FQ=-62.9905 GT:PL 0/0:0
    chr1 35399 . A . 61.2368 PASS DP=12;MQSB=0.95494;MQ0F=1;AF1=0;AC1=0;DP4=5,7,0,0;MQ=0;FQ=-62.9905 GT:PL 0/0:0
    chr1 35400 . A . 61.2368 PASS DP=12;MQSB=0.95494;MQ0F=1;AF1=0;AC1=0;DP4=5,7,0,0;MQ=0;FQ=-62.9905 GT:PL 0/0:0
    chr1 35401 . C . 61.2368 PASS DP=12;MQSB=0.95494;MQ0F=1;AF1=0;AC1=0;DP4=5,7,0,0;MQ=0;FQ=-62.9905 GT:PL 0/0:0

    As you can see, the chromosome position is continuous. I know that these are raw variant file but it contains 333,399,862 variants (almost as the number of bases). So, how can I filter this (there are so many false positives), i need filtration or should i do something at mpileup or bcftools call stage?

  • #2
    You want to call -cv to display only the variant positions, instead of call -c which shows all positions. You can see in your vcf it lists ref bases without an alt, so the positions in your example would all be removed.
    Providing nextRAD genotyping and PacBio sequencing services. http://snpsaurus.com

    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
    18 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 10:19 PM
    0 responses
    22 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 09:21 AM
    0 responses
    16 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-04-2024, 09:00 AM
    0 responses
    46 views
    0 likes
    Last Post seqadmin  
    Working...
    X