Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • samtools and IGV question

    I am new to samtools, IGV display and NGS in general. I have a two part question.

    Part 1-
    I would like to know how to interpret the output from "samtools mpileup" command. For example, in the attached txt file, column 5 has symbols like:
    1- "^]," what does this denote?
    2- "t" or "T" what is the difference between lower case and upper case?
    3- What is "-C"?
    4- why is "." or "," is used?
    and so on.

    Is there a comprehensive list where I can find all such information. Samtools FAQ has limited information. Any help is appreciated.
    (a text file is attached herein that I am trying to interpret)

    Part 2-
    Is there a samtools script or easily available script that can parse the information and tell us the allele frequency / total number of A,T,G, C at each mapped position. An icon displays frequency when mouse is moved over mapped position. It would be helpful if I can extract the information from the command line .. what IGV tools function / application would be used for this.

    Thanks in advance.
    Attached Files
    Last edited by fatakias; 01-16-2012, 07:51 AM. Reason: 2nd attachment

  • #2
    regarding part 1:

    1. a symbol `^' marks the start of a read segment which is a contiguous subsequence on the read separated by `N/S/H' CIGAR operations. The ASCII of the character following `^' minus 33 gives the mapping quality.
    2. T for a mismatch on the forward strand and t for a mismatch on the reverse strand
    3. -C -> deletion of C
    4. a dot stands for a match to the reference base on the forward strand, a comma for a match on the reverse strand

    Comment


    • #3
      This is the link to the page that explains pileup format:



      You can get allele counts from the DP4 values in the vcf, which is simpler than parsing the pileup. And its a much smaller file, too.

      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
      26 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 10:19 PM
      0 responses
      29 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 09:21 AM
      0 responses
      25 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