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  • Phuray
    Junior Member
    • Jul 2015
    • 1

    #1

    HELP!!!! How stable are Nextera Index primers?

    The problem has been solved, thanks a lot!
    Last edited by Phuray; 08-17-2015, 05:41 AM.
  • pmiguel
    Senior Member
    • Aug 2008
    • 2328

    #2
    They are probably fine, but if you don't want to take a chance, you could phone Illumina tech support and ask them. They might be willing to replace the primers.

    --
    Phillip

    Comment

    • GA-J
      Member
      • Jul 2015
      • 28

      #3
      I would simply test a few primers with index-PCR, and run on BA side by side.

      Comment

      • Simone78
        Senior Member
        • Oct 2010
        • 208

        #4
        they are quite robust, I have been freezing and thawing them probably 10 times or more and they were still fine. Unless you are using 50 ng of input DNA for your Nextera library prep, you have a huge excess of index primers anyway, so if part of them went bad, you are still safe.
        We use the Nextera XT kit, start from <500 pg for tagmentation and dilute the primers 1:5 to ame them last longer. And, even then, if we don´t do a bead clean up afterwards, we still see a huge peak of unused adaptors in the prep.

        Comment

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