Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Do my FASTQC results indicate sequencing bias?

    Hello,
    I used the Illumina Nextera Flex kit to sequence bacteria (WGS) and I got two warning that I usually do not get when using Nextera XT:

    1. overrepresented sequences in the first read (R1) but not the second read (R2).

    2. warning/error k-mer content

    Why do I get such errors? Does this indicate sequencing bias?
    Attached Files

  • #2
    This data must have been sequenced on NextSeq (or NovaSeq). That string of "G" is probably no-signal which is interpreted as G base.

    The bias at the beginning of the reads is likely due to the Nextera transposase sequence bias (similar to bias seen in "random priming") .

    If these are the only two "issues" then your data looks ok. Proceed with your normal analysis.

    Comment


    • #3
      Thank you for helping me!
      We use a MiseQ and it was the first time that we used Nextera Flex instead of XT. And it`s also the first time we got such a string of G.

      Beside these issues, I also got a warning for "per base sequence content". However, using Nextera XT always gives such issues.

      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 on Modified Bases...
        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
      39 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 10:19 PM
      0 responses
      41 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 09:21 AM
      0 responses
      35 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