10786357001

Rybonukleaza H (RNaza H)

from Escherichia coli H 560 pol A1

• Synonim(y): rnase h
• Kod UNSPSC: 12352204

Właściwości

• pochodzenie biologiczne: Escherichia coli ( H 560 pol A1)
• Poziom jakości: 100
• Próba: 100%
• Formularz: solution
• aktywność właściwa: ~40000 units/mg protein
• opakowanie: pkg of 100 U
• producent / nazwa handlowa: Roche
• metody: cDNA synthesis: suitable
• kolor: colorless
• optymalne pH: 7.5-9.1
• rozpuszczalność: water: miscible
• przydatność: suitable for molecular biology
• numer dostępu NCBI: NP_414750.1
• Zastosowanie: life science and biopharma
• obecność zanieczyszczeń: 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
• Warunki transportu: dry ice
• temp. przechowywania: −20°C (−15°C to −25°C)
• informacje o genach: Escherichia coli ... rnhA(946955)

Opis

Opis ogólny

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.

Zastosowanie

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)

Działania biochem./fizjol.

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.

Cechy i korzyści

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

Jakość

Absence of endonucleases, nicking activities, and ribonucleases.

Definicja jednostki

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

Uwaga dotycząca przygotowania

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

Przechowywanie i stabilność

Store at -15–-25 °C. (unopened)

Inne uwagi

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.

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Informacja o środkach bezpieczeństwa

• Kod klasy składowania: 12 - Non Combustible Liquids
• Klasa zagrożenia wodnego (WGK): WGK 1
• Temperatura zapłonu (°F): does not flash
• Temperatura zapłonu (°C): does not flash