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  • lee27
    Junior Member
    • Jul 2012
    • 3
    #1

    Ligating barcodes to amplicons

    Hi all,
    I am new to the forum and to next-gen in general. I am working on preparing a library of amplicons (20 different amplicons per individual sample - 96 to 180 samples) and I want to barcode the amplicons so that I can identify which individual they are from. Can barcodes/indices be directly ligated to the poly-A tail resulting from PCR or do I first need to blunt the end, apply a poly-A tail and then ligate? Thanks for your thoughts and patience!
    Tags: None
  • Zaag
    Senior Member
    • Nov 2009
    • 112
    #2
    Yes I would alwasy make it blunt end and perform the A-tailing. You could also design fusion primers with the adapter and MID so after PCR you can start emPCR without ligating.

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