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  • truseq DNA LS library preparation --- index adaptor issue

    I would like to make a pooled DNA solution and get it sequenced by hiseq 2000 Using just one lane.
    The pooled DNA solution contains 12 index adaptors from Truseq LS set B kit.
    When I see the protocol, Illumina recommends a certain combination of adaptors such as AD05 with AD12 (if pool 2 samples together). It looks like not all the index adaptors can be pooled together.
    I want to know if anyone has tried to sequence a DNA pool which contains all 12 index adaptors ? Can it be done? Or will cause serious problems?

  • #2
    Using specific adapter combinations is only an issue when you are doing "low-plex" pooling, meaning you are only pooling 2, 3 or 4 libraries in a lane. Beyond that you can add any other adapter(s) to the mix. You can pool all 12 from the TruSeq set B without a problem.

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    • #3
      Yes! i have tried all 12 index (TruSeq LT kit) and found no issue. All adaptor-index are compatible and samples de-multiplex easily. Platform used for sequencing was NexSeq 500.

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