Thermostable RNase H

Description:
Thermostable RNase H has an optimal activity above 65°C and can be used up to 95°C. The enzyme degrades RNA in a DNA:RNA hybrid, maximizing sensitivity and selectivity without affecting DNA or unhybridized RNA.

Application:
- High hybridization stringency
- Specific hydrolysis of RNA in a DNA:RNA hybrid
- Diagnostic assay of DNA sequences by isothermal probe amplification
- Mapping of mRNA structures

Source:
A recombinant protein purified from E. coli, cloned the gene encoding the Thermus thermophilus RNase H.

Unit Definition:
One unit of the enzyme 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 in 50 mM Tris-HCl(pH 7.5), 100 mM NaCl, and 10 mM 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.


10X RNase H Reaction Buffer
500 mM Tris-HCl(pH 7.5)
1 M NaCl
100 mM MgCl₂
pH 7.6 @ 25°C

Storage Buffer:
Supplied in 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.

Recommended Storage Condition: -20°C