NEBNext End Repair and dA-Tailing (E6115)


  1. 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 Volume: 9.0 μl
  2. Mix by pipetting and add to the 16 μl purified, double stranded cDNA.
  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 at 25°C
    20 minutes at 72°C
    Hold at 4°C