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  • Preparation of Adaptor Ligated DNA (E6270)

    Protocol

    1. Add the following to the microfuge tube:
        For 10 ng For 100 ng – 1 µg
      Sterile H2O 18 µl 3 µl
      T4 DNA Ligase Buffer for Ion Torrent 10 µl 10 µl
      Bst DNA Polymerase 1 µl 1 µl
      NEBNext DNA Library Adaptors for Ion Torrent 5 µl 20 µl
      T4 DNA Ligase 6 µl 6 µl
      Total volume 40 µl 40 µl


    2. The total volume in the microfuge tube should be 100 µl. Mix the contents by pipetting up and down several times.

    3. Incubate in a thermal cycler for 15 minutes at 25°C, followed by 5 minutes at 65°C.
    If performing size selection with beads, proceed directly to size selection using AMPure XP Beads. If using E-gel or agarose gel for size selection, cleanup adaptor ligated DNA before proceeding to size selection.