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Thermostable RNase H

Catalog # Concentration Size List Price Quantity Your Price
M0523S 5000 units/ml 250 units $236.00
$212.40
Catalog # Size List Price Your Price
M0523S 250 units $236.00
$212.40
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Thermostable RNase H specifically recognizes and cleaves the phosphodiester bonds of an RNA strand in an RNA-DNA hybrid while leaving the DNA strand intact. This thermostable nuclease exhibits the same enzymatic properties as E. coli RNase H, but is active at much higher temperatures.

  • Recombinant enzyme supplied with 10X Reaction Buffer
  • Does not digest single or double-stranded DNA
  • Active and stable above 37°C over a broad temperature range, with activity increasing with temperature
  • Requires magnesium chloride for activity
  • Inactivated with proteinase K treatment or addition of excess EDTA
Thermostable RNase H is an endoribonuclease that is functional at high temperatures and selectively hydrolyzes the phosphodiester bonds of an RNA strand in an RNA:DNA hybrid molecule, while leaving the DNA strand intact. In addition, Thermostable RNase H does not degrade single- or double-stranded RNA or DNA. While the Thermostable and E. coli RNase H homologs both exhibit these same endoribonucleolytic properties, the thermostable enzyme displays activity at higher temperatures with optimal activity above 65°C. These properties allow use in applications where higher assay temperatures are desired.
Product Source
An E. coli strain carrying a codon optimized plasmid encoding RNase H from the extreme thermophile Thermus thermophilus.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Thermostable RNase H M0523SVIAL -20 1 x 0.05 ml 5,000 units/ml
  RNase H Reaction Buffer B0297SVIAL -20 1 x 1.5 ml 10 X
Application Features
  • Higher stringency RNA structure mapping and site-specific RNA cleavage
  • Removal of poly(A) tails from mRNA hybridized to oligo(dT)
  • Removal of mRNA during second strand cDNA synthesis
  • Component of isothermal amplification methods

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to produce 1 nmol of ribonucleotides from 40 pmol of a fluorescently labeled 25 base pair RNA:DNA hybrid in a total reaction volume of 50 µl in 20 minutes at 50°C.

Reaction Conditions

1X RNase H Reaction Buffer
Incubate at ≥50°C

1X RNase H Reaction Buffer
50 mM Tris-HCl
75 mM KCl
3 mM MgCl2
10 mM DTT
(pH 8.3 @ 25°C)

Storage Buffer

50 mM Tris-HCl
100 mM NaCl
1 mM DTT
0.1 mM EDTA
0.1% Triton® X-100
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation

No

Notes
  • NOTE: The RNAse H Reaction Buffer contains MgCl2. When using Thermostable RNase H in the presence of the RNA:DNA hybrid of interest and other RNA species that are single-stranded (ie. total RNA) at elevated temperatures, it is advised to limit the reaction time and temperature to reduce metal-mediated breakdown of single-stranded RNA species.
  • Thermostable RNase H can be inactivated by treatment with proteinase K or addition of excess EDTA.
  • Optimal incubation temperature >65°C, up to 95°C.  Thermostable RNase H displays 3-4x more activity at 65°C compared to 37°C.
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
Legal And Disclaimer

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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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