10786357001

Ribonuclease H (RNase H)

from Escherichia coli H 560 pol A1

• Synonyme(s) : rnase h
• Code UNSPSC : 12352204

Propriétés

• Source biologique: Escherichia coli ( H 560 pol A1)
• Niveau de qualité: 100
• Pureté: 100%
• Forme: solution
• Activité spécifique: ~40000 units/mg protein
• Conditionnement: pkg of 100 U
• Fabricant/nom de marque: Roche
• Technique(s): cDNA synthesis: suitable
• Couleur: colorless
• pH optimal: 7.5-9.1
• Solubilité: water: miscible
• Adéquation: suitable for molecular biology
• Numéro d'accès NCBI: NP_414750.1
• Application(s): life science and biopharma
• Activité étrangère: RNase, none detected (up to 10 U with MS- II- RNA); endonuclease ~10 units, none detected (using lambda-DNA); nicking activity 10 units, none detected
• Conditions d'expédition: dry ice
• Température de stockage: −20°C (−15°C to −25°C)
• Informations sur le gène: Escherichia coli ... rnhA(946955)

Description

Description générale

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.

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.

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)

Actions biochimiques/physiologiques

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.

Caractéristiques et avantages

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

Qualité

Absence of endonucleases, nicking activities, and ribonucleases.

Définition de l'unité

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

Notes préparatoires

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

Stockage et stabilité

Store at -15–-25 °C. (unopened)

Autres remarques

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.

Informations sur la sécurité

• Code de la classe de stockage: 12 - Non Combustible Liquids
• Classe de danger pour l'eau (WGK): WGK 1
• Point d'éclair (°F): does not flash
• Point d'éclair (°C): does not flash