Hybridase Thermostable RNase H specifically degrades the RNA in a DNA:RNA hybrid, without affecting DNA or unhybridized RNA. In contrast to E. coli RNase H, which is rapidly inactivated at 55°C, Hybridase RNase H is active at high temperatures. It has optimal activity above 65°C and can be used at temperatures up to 95°C. The thermostability of the enzyme permits it to be used at temperatures that give the highest hybridisation stringency for specific DNA:RNA heteroduplexes, maximizing sensitivity and selectivity while minimizing background due to nonspecific hybridization.
Applications
Unit Definition: One unit of Hybridase RNase H results in the acid-solubilization of 1 nmol of polyadenylic acid in the presence of an equimolar concentration of polythymidylic acid in 20 minutes at 45°C under standard assay conditions.
Note: The unit assay is performed at 45°C because this is optimal for the T m of poly(dT):poly(A). The optimal temperature for many applications may be considerably higher.
Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 1.0 mM DTT, 0.1 mM EDTA, and 0.1% Triton® X-100.
Quality Control: Hybridase Thermostable RNase H is tested for RNA degradation in a RNA:DNA hybrid and for the absence of detectable exo- or endodeoxyribonuclease, and non-RNase H RNase activities.
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