Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Contamination between samples run on the same lane

    Hi colleagues,
    I found a strange effect when sequencing several samples on Miseq. The reads from each sample contain about ~0.1% of reads from other samples run on the same flowcell. This does not seem to be caused by improper recognition of barcodes - we did demultiplexing without allowing mismatches, and the indices are very different one from another (e.g. TAGCTT and AGTTCC).
    And this is certainly not contamination during library preparation - because I had one of the samples sequenced twice, with different "neighbors" and the contaminating reads were different.
    Have you ever seen something like that? What can be done to minimize this?

  • #2
    See if your observation is the same as recently mentioned here: http://seqanswers.com/forums/showthread.php?t=38697

    Comment


    • #3
      No, this is not a contamination between runs, if you mean this.

      Comment


      • #4
        Check the posts by vl80 and JackieBadger in the thread I linked above. They are about cross contamination between samples in the same run. That is what you are referring to right?

        Comment


        • #5
          Yeah, it looks like you're seeing what I've been calling index misassignment. It apparently stems from image alignment and signal bleed issues during index sequencing that causes a cluster to be assigned the wrong index sequence.

          Unfortunately, nothing can really be done about it, although dual indexing will cut down on the misassignment issue.

          Comment

          Latest Articles

          Collapse

          • seqadmin
            Recent Advances in Sequencing Analysis Tools
            by seqadmin


            The sequencing world is rapidly changing due to declining costs, enhanced accuracies, and the advent of newer, cutting-edge instruments. Equally important to these developments are improvements in sequencing analysis, a process that converts vast amounts of raw data into a comprehensible and meaningful form. This complex task requires expertise and the right analysis tools. In this article, we highlight the progress and innovation in sequencing analysis by reviewing several of the...
            05-06-2024, 07:48 AM
          • 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...
            04-22-2024, 07:01 AM

          ad_right_rmr

          Collapse

          News

          Collapse

          Topics Statistics Last Post
          Started by seqadmin, Today, 06:35 AM
          0 responses
          12 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, Yesterday, 02:46 PM
          0 responses
          18 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 05-07-2024, 06:57 AM
          0 responses
          17 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 05-06-2024, 07:17 AM
          0 responses
          18 views
          0 likes
          Last Post seqadmin  
          Working...
          X