During RNA/nuclei isolations we usually have DTT or Beta mercaptoethanol in our buffers which basically reduces the disulfide bonds of the RNases thereby inhibiting them. My question is - since these RNAse Inhibitors are also proteins and might have cystine residues which then forms disulfide bonds, does the DTT or Beta-ME degrade these inhibitors as well?
Really good question! I don't know exactly why, but most RNase inhibitors actually require reducing conditions to work properly. Adding expensive RNase inhibitors to buffers that do not contain DTT is probably one of the most common mistakes I see in RNA protocol design. The only RNase inhibitor which works in non-reducing conditions is Superase.in, which is one of the most expensive RNase inhibitors on the market - it's situationally very useful for experiments that are incompatible with DTT, but it also works fine with very high concentrations of DTT present. Murine RNase inhibitor from NEB works with lower DTT concentrations than most other inhibitors, and is also relatively cheap.
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