Cleavage of an RNA strand in an RNA-DNA hybrid with RNase H (NEB #M0297)

RNase H specifically recognizes and cleaves the phosphodiester bonds of an RNA strand in an RNA-DNA hybrid while leaving the DNA strand intact. The RNase H will only cleave the RNA in an RNA:DNA hybrid.

1. Set-up a typical reaction as follows:

   COMPONENTS	100 μl REACTION
   RNA:DNA duplex	2 µg
   RNase H Reaction Buffer(10X)	10 µl (1X)
   RNase H	1 µl (5 units)
   Nuclease-free H2O (NEB#B1500S)	up to 100 µl

2. Incubate at 37°C for 20 minutes.
3. The reaction can be stopped with addition of 1 µl of 0.5 M EDTA.

* Note: If targeting an RNA in a pool of RNA, we recommend titration of the enzyme to the intended substrate. Optimization may be required based on sequence, length and the type of reaction.