FAQ: Can NEBNext® Magnesium RNA Fragmentation Module be used instead of the NEBNext® RNase III RNA Fragmentation Module?

Yes, however the ends created by the two kits are different. If 5' phosphate and 3' hydroxyl termini are required, end repair can be performed as follows:

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

  1. Incubate at 37°C for 30 minutes.
  2. Add 2ul 10 mM ATP and continue incubation at 37°C for 30 minutes.