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  • End Repair of DNA Protocol (E6270)

    Overview

    Starting Material: 10 ng–1 μg of Fragmented DNA.

    Note: For use with the Ion Xpress™ Barcode Adapters 1-16 Kit, a minimum of 100 ng starting material is recommended. Lower amounts may cause adaptor concatamerization.

    Protocol

    1. Mix the following components in a sterile microfuge tube on ice:
      Fragmented DNA   1–51 μl
      NEBNext End Repair Reaction Buffer   6 μl
      NEBNext End Repair Enzyme Mix   3 μl
      Sterile H2O   variable
      -------------------------------------------------------------------------
      Total volume   60 μl

    2. Incubate in a thermal cycler for 20 minutes at 25°C, followed by 10 minutes at 70°C.

    3. Pulse spin the microfuge tube and return to ice.