NEBNext End Repair and dA-Tailing (E6080)
- 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
- Mix by pipetting and add to the 16 μl fragmented DNA sample.
- 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 @ 25°C
20 minutes @ 72°C
Hold at 4°C