Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Poor per tile sequence quality

    We recently performed a couple HiSeq runs, and I'm quite new to NGS analysis.

    I'm attaching a representation of the results from FastQC for most of the samples. I'm wondering if anyone knows what might account for the poor sequencing quality.

    Thank you!

    Click image for larger version

Name:	seq_qual.png
Views:	1
Size:	36.5 KB
ID:	309266

    Click image for larger version

Name:	per_tile.png
Views:	1
Size:	35.1 KB
ID:	309267

  • #2
    I'm guessing it's poor flow in certain tiles, which yields worse quality with every cycle as the read positions in the colony become increasingly out-of-sync (some percent fail to advance with each cycle).

    Comment


    • #3
      Thanks Brian! Would the poor flow be due to overloading of sample into the flow cell? Or, are there are reasons for poor flow in specific tiles? (ie: machine-specific reasons or others?)

      Thank you!

      Comment


      • #4
        Overloading shouldn't have any impact on flow. Flow problems are typically caused by machine-specific fluidics issues (clogged nozzles, etc), I believe.

        Comment


        • #5
          Oh, I see. Thanks so much! If anyone else has thoughts on the results, please let me know!

          Thank you!

          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
          30 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-10-2024, 10:19 PM
          0 responses
          32 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-10-2024, 09:21 AM
          0 responses
          28 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 04-04-2024, 09:00 AM
          0 responses
          53 views
          0 likes
          Last Post seqadmin  
          Working...
          X