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RNase H

Ribonuclease H (RNase H) is an endoribonuclease that specifically degrades the RNA strand in RNA-DNA hybrids.


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Description
Specifications
Supporting Data
References
Resources

Thermo Scientific Ribonuclease H (RNase H) specifically degrades the RNA strand in RNA-DNA hybrids (see Figure 1 in Supporting Data). It does not hydrolyze the phosphodiester bonds within single-stranded and double-stranded DNA and RNA.

Applications

Removal of mRNA prior to synthesis of second strand cDNA (see Reference 1)
RT-PCR and qRT-PCR: removal of RNA after first strand cDNA synthesis
Removal of the poly(A) sequences of mRNA after hybridization with oligo(dT) (see Reference 2)
Site-specific cleavage of RNA (see Reference 3)
Studies of in vitro polyadenylation reaction products (see Reference 4)


Storage Condition	-20 C
Hazardous	No
10X Reaction Buffer	200 mM Tris-HCl (pH 7.8), 400 mM KCl, 80 mM MgCl₂, 10 mM DTT
Concentration	5 U/µL
Definition of Activity Unit	One unit of the enzyme catalyzes the formation of 1 nmol of acid soluble products in 20 min at 37°C. Enzyme activity is assayed in the following mixture: 20 mM Tris-HCl (pH 7.8), 40 mM KCl, 8 mM MgCl₂, 1 mM DTT, 24 µM [³H]-poly(A)·poly(dT), 0.03 mg/mL BSA, 4% (v/v) glycerol.
Inactivation	Inactivated by heating at 65°C for 10 min.
Inhibition	Inhibitors: metal chelators, SH-blocking reagents.
Molecular Weight	18.4 kDa monomer
Quality Control	The absence of endo-, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests.
Source	E.coli MRE-600 cells
Storage Buffer	The enzyme is supplied in: 25 mM HEPES-KOH (pH 8.0), 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.1 mg/mL BSA, and 50% (v/v) glycerol.