NEBNext End Repair and dA-Tailing (E6115)

Protocol

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