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 r1.1–r3.1 and rCutSmart® Buffer:

Buffer 37°C 50°C 65°C >65°C
RNase H Buffer +++ +++ +++ +++
NEBuffer 1 ++ ++ ++ ++
NEBuffer 2; +++ +++ ++ ++
NEBuffer 3 +++ +++ +++ +
NEBuffer 4 ++ +++ +++ +++
NEBuffer r1.1 ++ ++ +++ -
NEBuffer r2.1 ++ ++ ++ ++
NEBuffer r3.1 +++ +++ +++ -
rCutSmart Buffer ++  ++ ++ +

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