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  • PCR primers+adaptors

    Hi, I am working for my Ms thesis on a metagenetic study of the meiofauna and I am going to amplify the 18S region, then sequencing with 454. My question is if I should reshape primers with te A and B adaptors, and the MID tags before amplify my samples.
    I read the B adaptor has a biotina than should not be a normal primer. In every works that I read they always amplify 18s with adaptors and mid tags referring as normal primers. Does anyone can help me?

    All the best

  • #2
    You shouldn't need modified primers for what you want to do. Just append the adapter sequences and MIDs to the amplification primers before ordering. The MID only goes on the A primer, so you'll need a different one for each sample plus a common reverse (same for every sample) with the B adapter sequence appended.
    There's a tech note somewhere on the my454 website with the sequences and design strategy.

    Comment


    • #3
      Thanks a lot TonyBrooks!

      Comment

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