Ribonuclease H ( RNase H ) is an endo-ribonuclease, which can specifically degrade the phosphodiester bond of RNA hybridized to the DNA strand, so it can degrade the RNA strand in the RNA / DNA hybrid strand. This enzyme cannot digest single- or double-stranded DNA. This RNase H enzyme is widely used to remove mRNA poly (A) hybridized to poly (dT) or to remove mRNA during second-strand synthesis of cDNA.
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Cat. No.: RNH-100 (for 250U)
Cat. No.: RNH-1k (for 1000U)
Cat. No.: RNH-5k (for 5000U)
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.
It specifically digests RNA in DNA-RNA hybrid chains and is commonly used in the synthesis of the second strand of cDNA.
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.
Purified from recombinant E. coli strains carrying the gene encoding RNase H from Escherichia coli.
Approximately 18.4 kDa (monomer).
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.
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.
When using the enzyme, it should be stored in an icebox or on an ice bath. After use, it should be immediately stored at -20°C.
This product is intended for scientific research purposes by professionals only. It is not to be used for clinical diagnosis or treatment, and should not be used in food or drugs. It should not be stored in a regular household.
For your safety and health, please wear a lab coat and disposable gloves when handling.