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).