Hybridase-Thermostable RNase H, 500 U @ 5 U/uL

Specifically degrade the RNA in a DNA:RNA hybrid, without affecting DNA or unhybridized RNA, at higher reaction temperatures

• Optimal activity above 65°C and maintains activity as high as 95°C
• Highly specific for RNA in a RNA:DNA hybrid and will not digest free RNA or DNA
• Maximises digestion sensitivity and selectivity while minimising background due to nonspecific hybridization

Applications

• High-stringency hybrid selection.
• Diagnostic assays of specific target DNA sequences by isothermal probe amplification.^1,2
• Transcription-based amplification methods (e.g., NASBA® method).^3,4
• High-stringency mapping of mRNA structure.
• Applications that require specific hydrolysis of the RNA in a DNA:RNA hybrid.

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 hybridization stringency for specific DNA:RNA heteroduplexes, maximizing sensitivity and selectivity while minimising background due to nonspecific hybridization.

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.

References

1. Duck, P. et al. (1990) BioTechniques 9, 142.
2. Bekkaoui, F. et al. (1996) BioTechniques 20, 240.
3. Guatelli, J.C. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1874.
4. Fahy, E. et al. (1991) PCR Methods and Applications 1, 25.