RNase H

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recombinant unique buffer incubation temp heat inactivation
Catalog #SizeConcentrationPriceQtyAdd to Cart
M0297S250 units5,000 units/ml$67.00Add to Cart
M0297L1,250 units5,000 units/ml$268.00Add to Cart
  
Categories:
cDNA Synthesis & Reverse Transcriptases,
cDNA Synthesis & Reverse Transcriptases,
RNases Products
Applications:
cDNA Synthesis & RT-PCR,
RT-PCR & cDNA Synthesis,
RT-PCR & cDNA Synthesis

Description

RNase H (Ribonuclease H ) is an endoribonuclease that specifically hydrolyzes the phosphodiester bonds of RNA which is hybridized to DNA. This enzyme does not digest single or double-stranded DNA.

Product Source

An E. coli strain that carries the cloned RNase H gene (rnh) from Escherichia coli.

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
RNase H Reaction Buffer-2010X

Properties and Usage

Unit Definition

One unit is defined as the amount of enzyme required to produce 1 nmol of ribonucleotides from 20 picomoles of a fluorescently labelled 50 base pair RNA-DNA hybrid in a total reaction volume of 50 µl in 20 minutes at 37°C.

Reaction Conditions

1X RNase H Reaction Buffer
Incubate at 37°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 Temperature

-20°C

Storage Conditions

50 mM KCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
200 μg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 min

References

  1. Davis, R. et al. (1988). Cell Biol. 8, 4745-4755.
  2. Gubbler, U. and Hoffman, B.J. (1983). Gene. 25, 263-269.
  3. Donnis-Keller, H. (1979). Nucl. Acids Res. 7, 179.
  4. Goodwin, E.C. and Rottman, F.M. (1992). Nucl. Acids Res. 20, 916.

FAQs

  1. Is RNase H active in NEBuffer™ 1-4?
  2. Is DNA Polymerase I active in RNase H buffer?
  3. Which side of the RNA base does RNase H cut?
  4. What is the activity of RNase H at 30°C?

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Interactive Tools

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • Endonuclease Activity (Nicking):
    The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
  • RNase Activity (2 Hour Digestion):
    The product is tested in a reaction containing a RNA substrate.  After incubation for 2 hour there is no detectable degradation of the RNA substrate as determined by gel electrophoresis.
  • Single-Stranded DNase Activity (Radioactivity Release):
    The product is tested in reaction containing radiolabeled single-stranded DNA. After incubation the exonuclease activity is determined by the % release of radioactive nucleotides.

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.