Ribonuclease H (RNase H)

Uses

Ribonuclease H (RNase H) has been used for:

• In vivo RNA-primed initiation of DNA synthesis
• Elimination of mRNA during second-strand cDNA synthesis
• Site-specific cleavage of RNA
• Detection of RNA:DNA regions in double-stranded DNA of natural origin
• Removal of poly (A) sequences of mRNA if oligo (dT) is present
• RNA extraction and quantitative reverse transcriptase polymerase chain reaction (RT-PCR)

General Description

Nonspecific endoribonuclease that specifically cleaves RNA in RNA:DNA hybrids. A minimum of four continuous base pairs (RNA:DNA) is required for activity. RNase H cleaves RNA to release 5′-oligoribonucleotides.

Source: E. coli H560 pol A1
Storage Buffer: 25 mM Tris-HCl, 50 mM KCl, 1 mM dithiothreitol, 0.1 mM EDTA, 50% glycerol (v/v), pH 8.0 (+4°C)
Volume Activity: 1 x 103 U/ml assayed according to Hillenbrand & Staudenbauer.

Biochem/physiol Actions

Ribonuclease H (RNase H) specifically cleaves RNA in RNA:DNA hybrids. A minimum of four continuous base pairs (RNA:DNA) is required for activity. RNase H cleaves RNA to release 5′-oligoribonucleotides. RNase H is associated with nucleic acid immunity. Using RNase H for degrading mRNA results in 80% depletion of mRNA and protein expression. RNase H recognizes the start codon and the 3′ and 5′ untranslated regions. This enzyme participates in DNA replication.