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  • seqgirl123
    Member
    • Oct 2008
    • 78

    Illumina sequences with Kapa kit

    Has anyone used Illumina Truseq adapters and the PPC primer cocktail with the Kapa library preparation kit (Illumina compatible version)? The Kapa kit allows the user to use outside adapter and PCR primer primers, this is the reason why I'm asking, if anyone has tried it and how it worked?

    Also I am asking because I want to try Kapa's HiFi polymerase. But since I plan to use Truseq adapters, I am at a loss to how to synthesize Truseq PCR primers to use along with Kapa's polymerase, and since Illlumina doesn't publish their PCR primer sequences anymore, I don't know what other PCR primers to use (besides qPCR primers) other than the PPC primer cocktail. Has anyone explored this?
  • luc
    Senior Member
    • Dec 2010
    • 469

    #2
    I did try the Kapa kit about a year ago. It worked very well. The one drawback is that it requires the old-style cleanup step between A-tailing and adapter ligation; with the Illumina kit or home-brewed protocols you can carry out the A-tailing in ligation buffer and thus skip one of the cleanup steps. The one kit, that I used was also a bit low on reagents - might have been an accident.
    For the Kapa Hifi mastermix we are using an annealing temp of 63 degrees instead of 65; otherwise everything is standard.

    Info on truseq adapters and primers is available here:
    Last edited by luc; 03-29-2013, 09:44 AM.

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