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  • RNase H

    Product Update:  The product storage buffer has been updated to a standard storage buffer.
    Did you know this product can be customized or purchased in larger volumes? Submit an inquiry to find out more.
    recombinant unique buffer incubation temp heat inactivation
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    M0297S250 units5,000 units/ml$66.00Add to Cart
    M0297L1,250 units5,000 units/ml$264.00Add to Cart
      
    Categories:
    cDNA Synthesis & Reverse Transcriptases,
    RNases Products
    Applications:
    cDNA Synthesis & RT-PCR,
    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.
    • 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.