NEBNext End Repair and dA-Tailing (E6115)
Protocol
- 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
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Total Volume: 9.0 μl - Mix by pipetting and add to the 16 μl purified, double stranded cDNA.
- Vortex briefly to mix, followed by a quick spin to collect all liquid from the sides of the tube.
- 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