Ribonuclease H (RNase H)
ROCHE/10786357001 - from Escherichia coli H 560 pol A1
Synonym: rnase h
Product Type: Chemical
| application(s) | life science and biopharma |
| assay | 100% |
| biological source | Escherichia coli ( |
| color | colorless |
| foreign activity | endonuclease ~10 units, none detected (using lambda-DNA) |
| nicking activity 10 units, none detected | |
| RNase, none detected (up to 10 U with MS- II- RNA) | |
| form | solution |
| manufacturer/tradename | Roche |
| NCBI accession no. | NP_414750.1   |
| optimum pH | 7.5-9.1 |
| packaging | pkg of 100 U |
| Quality Level | 100 |
| shipped in | dry ice |
| solubility | water: miscible |
| specific activity | ~40000 units/mg protein |
| storage temp. | −20°C (−15°C to −25°C) |
| suitability | suitable for molecular biology |
| technique(s) | cDNA synthesis: suitable |
| Application: | 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) |
| 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 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. |
| General description: | 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 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| Purity | 100% |
| activity | specific activity: ~40000 units/mg protein |
| Storage Temp. | −20°C (−15°C to −25°C) |
| UNSPSC | 12352204 |