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Hybridase Thermostable RNase H

Hybridase Thermostable RNase H

Specifically degrade the RNA in a DNA:RNA hybrid, without affecting DNA or unhybridised RNA, at higher reaction temperatures.
  • Specifically degrade the RNA in a DNA:RNA hybrid, without affecting DNA or unhybridised 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 hybridisation
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Product information

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

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

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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