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  • NEBNext End Repair and dA-Tailing (E6080)

    Protocol

    1. Starting Material: 0.5 μg of DNA Fragmented to 100–1000 bp in 16 μl of TE.

      In a 1.7 ml micro-centrifuge tube add:
      NEBuffer 2 (10X) 2.5 μl
      ATP 2.5 μl
      dNTP Mix 1.0 μl
      T4 DNA Polymerase 1.0 μl
      PNK 1.0 μl
      Taq DNA Polymerase 1.0 μl
      Total 9.0 μl


    2. Mix by pipetting and add to the 16 μl fragmented DNA sample.

    3. Vortex briefly to mix, followed by a quick spin to collect all liquid from the sides of the tube.

    4. In a thermocycler, with the heated lid on, run the following program: 
      20 minutes @ 25°C 
      20 minutes @ 72°C 
      Hold at 4°C