Add the following to your fragmented and purified sample:
Murine RNase Inhibitor (NEB# M0314)
0.5 ul T4 PNK Buffer , 10X(NEB# B0201)
2 ul T4 PNK Enzyme (NEB# M0201)
1 ul Nuclease-Free Water 2.5 ul
Total volume 18 ul
- Incubate at 37°C for 30 minutes.
- Add 2ul 10 mM ATP and continue incubation at 37°C for 30 minutes.