FAQ: Is Thermostable RNase H active in other buffers?

Thermostable RNase H requires Mg2+ for activity, therefore buffers containing MgCl2 may be suitable for Thermostable RNase H reactions.  The optimal reaction buffer is 1X RNase H Reaction Buffer, but we have tested NEBuffers 1–4, NEBuffers 1.1–3.1 and CutSmart® Buffer:

Buffer 37°C 50°C 65°C >65°C
RNase H Buffer +++ +++ +++ +++
NEBuffer 1 ++ ++ ++ ++
NEBuffer 2; +++ +++ ++ ++
NEBuffer 3 +++ +++ +++ +
NEBuffer 4 ++ +++ +++ +++
NEBuffer 1.1 ++ ++ +++ -
NEBuffer 2.1 ++ ++ ++ ++
NEBuffer 3.1 +++ +++ +++ -
CutSmart ++  ++ ++ +