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10786357001

Roche

Ribonuclease H (RNase H)

from Escherichia coli H 560 pol A1

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Synonym(s):
rnase h
UNSPSC Code:
12352204

biological source

Escherichia coli ( H 560 pol A1)

Quality Level

assay

100%

form

solution

specific activity

~40000 units/mg protein

packaging

pkg of 100 U

manufacturer/tradename

Roche

technique(s)

cDNA synthesis: suitable

color

colorless

optimum pH

7.5-9.1

solubility

water: miscible

suitability

suitable for molecular biology

NCBI accession no.

application(s)

life science and biopharma

foreign activity

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

shipped in

dry ice

storage temp.

−20°C (−15°C to −25°C)

Gene Information

Escherichia coli ... rnhA(946955)

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.
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)

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.

Quality

Absence of endonucleases, nicking activities, and ribonucleases.

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

Preparation Note

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

Storage and Stability

Store at -15–-25 °C. (unopened)

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.

wgk_germany

WGK 1

flash_point_f

does not flash

flash_point_c

does not flash


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