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  • NEBNext End Prep (E7442)

    Protocol

    Starting Material: 5 ng–1 μg fragmented DNA

    1. Mix the following components in a sterile, nuclease-free tube:
      End Prep Enzyme Mix   3.0 μl
      End Repair Reaction Buffer (10X)   6.5 μl
      Fragmented DNA   55.5 μl
      -------------------------------------------------------------------
      Total volume   65 μl
    2.  Mix by pipetting, followed by a quick spin to collect all liquid from the sides of the tube.
    3. Place in a thermocycler, with the heated lid on, and run the following program:
      30 minutes @ 20°C
      30 minutes @ 65°C
      Hold at 4°C
    4. Proceed directly to NEBNext Ultra Ligation Module (NEB #E7445).