10786357001

Ribonuclease H (RNase H)

from Escherichia coli H 560 pol A1

• Sinonimo/i: rnase h
• Codice UNSPSC: 12352204

Proprietà

• Origine biologica: Escherichia coli ( H 560 pol A1)
• Livello qualitativo: 100
• Saggio: 100%
• Forma fisica: solution
• Attività specifica: ~40000 units/mg protein
• Confezionamento: pkg of 100 U
• Produttore/marchio commerciale: Roche
• tecniche: cDNA synthesis: suitable
• Colore: colorless
• pH ottimale: 7.5-9.1
• Solubilità: water: miscible
• Compatibilità: suitable for molecular biology
• N° accesso NCBI: NP_414750.1
• applicazioni: life science and biopharma
• Attività estranea: 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
• Condizioni di spedizione: dry ice
• Temperatura di conservazione: −20°C (−15°C to −25°C)
• Informazioni sul gene: Escherichia coli ... rnhA(946955)

Descrizione

Descrizione generale

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.

Applicazioni

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)

Azioni biochim/fisiol

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.

Caratteristiche e vantaggi

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

Qualità

Absence of endonucleases, nicking activities, and ribonucleases.

Definizione di unità

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

Nota sulla preparazione

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

Stoccaggio e stabilità

Store at -15–-25 °C. (unopened)

Altre note

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.

Informazioni sulla sicurezza

• Codice della classe di stoccaggio: 12 - Non Combustible Liquids
• Classe di pericolosità dell'acqua (WGK): WGK 1
• Punto d’infiammabilità (°F): does not flash
• Punto d’infiammabilità (°C): does not flash