Not available outside of the UK & Ireland.

Application

Ribonuclease H (RNase H) has been used for:In vivo RNA-primed initiation of DNA synthesisElimination of mRNA during second-strand cDNA synthesisSite-specific cleavage of RNADetection of RNA:DNA regions in double-stranded DNA of natural originRemoval of poly (A) sequences of mRNA if oligo (dT) is presentRNA extraction and quantitative reverse transcriptase polymerase chain reaction (RT-PCR)

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.

Features and Benefits

Eliminate potential sources of PCR errors. Increase accessibility of primers during subsequent 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 A1Storage 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.

Ribonuclease H (RNase H) is a nonspecific endoribonuclease, localized to the nucleus and cytoplasm. It is ubiquitously found and widely present among many organisms including viruses and human.

Other Notes

For life science research only. Not for use in diagnostic procedures. Using RNase H after the cDNA synthesis step can increase the sensitivity of a two-step RT-PCR assay.

Preparation Note

Activator: The enzyme has its maximal activity in presence of SH-reagents

Quality

Absence of endonucleases, nicking activities, and ribonucleases.

Storage and Stability

Store at -15–-25 °C. (unopened)

Unit Definition

RNase H is assayed according to Hillenbrand and Staudenbauer. One unit of RNase H is the amount of enzyme which produces 1 nmol acid-soluble ribonucleotides from[³H] poly(A) x poly(dT) in 20 minutes at +37 °C under the stated assay conditions.Volume Activity: Approximately 1 U/µl