RNase H, Tth

Description
Thermostable RNase H specifically degrades the RNA in a DNA:RNA hybrid, without affecting DNA or unhybridized RNA. This thermostable nuclease exhibits the same enzymatic properties as E. coli RNase H, but is active at much higher 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 minimizing background due to nonspecific hybridization.

Applications

• Removal of poly(A) tails of mRNA hybridized to poly(dT)
• Removal of mRNA during second strand cDNA synthesis

Source:
A recombinant E. coli strain carrying the RNAse H gene from the thermophilic eubacterium Thermus thermophilus HB8.

Storage Buffer:

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 and 50% glycerol.

10x RNAse H Buffer:
500 mM Tris-HCl, 750 mM KCl, 30 mM MgCl_2, 100 mM DTT, pH 8.3 @ 25°C

Unit Definition:
One unit results in the acid-solubilization of 1 nmole of [3H]-polyadenylic
acid in the presence of an equimolar concentration of polythymidylic acid in 20 minutes at 45°C.

Recommended Storage Condition: -20°C