Epicentre's® patented* Hybridase™ Thermostable RNase H specifically degrades only RNA in an DNA:RNA hybrid, without affecting DNA or unhybridised 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, maximising sensitivity and selectivity while minimising background due to nonspecific hybridisation.
One unit results in the acid-solubilisation of 1nmole of 3H-polyadenylic acid in the presence of an equimolar concentration of polythymidylic acid in 20 minutes at 45°C in 50mM Tris-HCl, pH 7.5, 100mM NaCl, and 10mM MgCl2.
Note: The unit assay is performed at 45°C because this is optimal for the Tm of poly(dT):poly(A). The optimal temperature for many applications may be considerably higher.
50% glycerol containing 50mMTris-HCl, pH 7.5, 0.1M NaCl, 1.0 mM DTT, 0.1mM EDTA, and 0.1% Triton® X-100.
Hybridase Thermostable RNase H is tested for RNA degradation in an RNA:DNA hybrid and for the absence of detectable DNA exo- or endonuclease, and non-RNase H RNase activities.
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