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  • Custom primers for NextSeq: will they work on MiSeq and HiSeq?

    I am wondering whether my experiment (which was originally designed to work on a NextSeq) will work on MiSeq and HiSeq.

    It is a library prepared with MIP technique and the PCR primers are

    CAAGCAGAAGACGGCATACGAGATNNNNNNNNACACGCACGATCCGACGGTAGTGT
    AATGATACGGCGACCACCGAGATCTACACATACGAGATCCGTAATCGGGAAGCTGAAG

    My sequencing primers:

    FOR CATACGAGATCCGTAATCGGGAAGCTGAAG C
    REV ACACGCACGATCCGACGGTAGTGT A
    INDX ACACTACCGTCGGATCGTGCGTGT A

    I would be grateful for a quick response

    Anna

  • #2
    We use custom primers and run libraries on NextSeq and HiSeq. MiSeq has a higher temperature for binding so a given primer may not work on MiSeq.
    Providing nextRAD genotyping and PacBio sequencing services. http://snpsaurus.com

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    • #3
      Thanks for your reply!
      OK, now, how do I check that? What are the boundaries for Tm for MiSeq, NextSeq and HiSeq?

      Comment

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