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  • MLog
    Member
    • Jan 2010
    • 36

    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?
  • GenoMax
    Senior Member
    • Feb 2008
    • 7142

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

    Comment

    • MLog
      Member
      • Jan 2010
      • 36

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

      Comment

      • GenoMax
        Senior Member
        • Feb 2008
        • 7142

        #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

        • mcnelson.phd
          Senior Member
          • Jul 2011
          • 162

          #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

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