E. coli; cRNAse H
Ribonuclease H specifically degrades only the RNA strand in RNA-DNA hybrids. It does not hydrolyze the phosphodiester bonds within single-stranded or double-stranded DNA and RNA.
Unit Definition
One unit catalyzes the formation of 1nmol of acid soluble products in 20min at 37ºC. Enzyme activity is assayed in the following mixture: 20mM Tris-HCl (H 7.8), 40 mM KCl, 8mM MgCl2, 1mM DTT, 24uM [3H]-POLY(A)-POLY(dT), 0.03mg/ml BSA AND 4% Glycerol.
10X Reaction buffer (Included)
R2021A Supplied as a liquid in 200mM Tris HCl, pH 8.0, 400mM KCl, 80mM MgCl2, 10mM DTT.
Applications
Suitable for use in Removal of mRNA prior to the synthesis of the 2nd strand of cDNA, Removal of mRNA poly(A) sequences after hybridization with oligo(dT), Site-specific cleavage of RNA and Studies of in vitro polyadenylation rection products.
Inhibition and Inactivation
-Inhibitors: metal chelators, SH-blocking reagents. -Inactivated by heating at 65ºC for 10min.
Endodeoxyribonuclease Assay
No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 10 units of Ribonuclease H with 1ug of pBR322 DNA in 50ul of reaction buffer for 1 hour at 37°C.
Exodeoxyribonuclease Assay
0% of the total radioactivity was released into trichloroacetic acid-soluble fraction after incubation of 10 units of Ribonuclease H with 1ug of sonicated E.coli [3H]-DNA in 50ul of reaction buffer for 1 hour at 37°C.
Ribonuclease Assay
0% of the total radioactivity was released into trichloroacetic acid-soluble fraction after incubation of 10 units of Ribonuclease H with 1ug of [3H]-RNA in 50ul of reaction buffer for 1 hour at 37°C.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Source
E. coli MRE-600 cells
Form
Supplied as a liquid in 25mM HEPES-KOH, pH 8.0, 50mM KCl, 1mM DTT, 1mM EDTA, 0.1mg/ml BSA, and 50% glycerol.
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
Form
Supplied as a liquid in 25mM HEPES-KOH, pH 8.0, 50mM KCl, 1mM DTT, 1mM EDTA, 0.1mg/ml BSA, and 50% glycerol.
TDS
NCBI
UniProt
Tech Support
Customer Service
