Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • problem with R2 reads from Miseq

    Dear, I have a problem in Miseq data. I have paired 2x150bp reads. I have identified some false positive (repetitive in different samples) due to short mapped reads. In fastq files (both R1 and R2) I have the reads complete, but sometimes only little fragments (about 35bp) of reads from R2.fastq map to genome, as PCR duplicates .
    The related reads in always in *R2.fastq, and if I look into bam file this is "Second in pair".

    Anyone has any idea? Is it an experimental problem or a bioinformatic one?
    Thanks
    Attached Files

  • #2
    problem with R2 reads from Miseq

    What software was used to align the reads, and were the reads trimmed before alignment?

    Comment


    • #3
      I use bwa, samtools rmdup to remove duplicates (not -S option), gatk for local realignment and recalibration.
      The reads before alignment are trimmed randomly. In the figure you can see three reads in R2 fastq file and in yellow the mapped short read. I don't know what happens to the rest of bp.
      Attached Files

      Comment

      Latest Articles

      Collapse

      • seqadmin
        Essential Discoveries and Tools in Epitranscriptomics
        by seqadmin




        The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist...
        Yesterday, 07:01 AM
      • 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

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by seqadmin, 04-11-2024, 12:08 PM
      0 responses
      58 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 10:19 PM
      0 responses
      54 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 09:21 AM
      0 responses
      46 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-04-2024, 09:00 AM
      0 responses
      55 views
      0 likes
      Last Post seqadmin  
      Working...
      X