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Cat. No.: RNH-100 (for 250U)
Cat. No.: RNH-1k (for 1000U)
Cat. No.: RNH-5k (for 5000U)
Description
RNase H, also known as Ribonuclease H, is a ribonucleic acid endonuclease obtained through expression and purification from Escherichia coli. It specifically hydrolyzes the RNA in DNA-RNA hybrid chains. RNase H cannot hydrolyze the phosphodiester bonds in single-stranded or double-stranded DNA or RNA, which means it cannot digest single-stranded or double-stranded DNA or RNA. RNase H is widely used to digest and degrade RNA in DNA-RNA hybrid double-strands formed after the synthesis of the second strand of cDNA, or to remove mRNA poly(A) hybridized to oligo (dT), among other applications.
Features
It specifically digests RNA in DNA-RNA hybrid chains and is commonly used in the synthesis of the second strand of cDNA.
Applications
Removal of RNA from DNA-RNA hybrid chains, such as pre-removal of RNA before the synthesis of the second strand of cDNA; removal of RNA after the synthesis of the first strand of cDNA in RT-PCR/RT-qPCR experiments; site-specific cleavage of RNA using complementary DNA sequences; removal of poly(A) from mRNA hybridized to poly(dT); studies of products in vitro polyadenylation reactions; identification of DNA-RNA hybrid double-strands.
Source
Purified from recombinant E. coli strains carrying the gene encoding RNase H from Escherichia coli.
Molecular weight
Approximately 18.4 kDa (monomer).
Activity definition
One unit is defined as the amount of enzyme required to produce 1 nmol of ribonucleotides from 20 picomoles of a fluorescently labeled 50 base pair RNA-DNA hybrid in a total reaction volume of 50 µl in 20 minutes at 37°C.
Purity
Free of DNA endonucleases and exonucleases, and other ribonucleases.
Enzyme storage solution
10 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 200 μg/ml BSA, 50% (v/v) glycerol, (pH 7.4 @ 25°C).
Reaction Buffer (10X)
500 mM Tris-HCl, 750 mM KCl, 30 mM MgCl2, 100 mM DTT, (pH 8.3 @ 25°C).
Inactivation or inhibition
RNase H can be inactivated by heating at 65°C for 10 minutes. Metal ion chelators and thiol-blocking agents can inhibit RNase H activity. Addition of EDTA to a final concentration of at least 5 mM can inhibit the enzymatic activity of RNase H.
Precautions
Storage
Store at -20°C, valid for 2 years.
Related:
Only for research and not intended for treatment of humans or animals
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