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  • DNase I (RNase-free)

    Description

    DNase I, (RNase-free) is an endonuclease that nonspecifically cleaves DNA to release di-, tri- and oligonucleotide products with 5´-phosphorylated and 3´-hydroxylated ends (1,2). DNase I acts on single- and double-stranded DNA, chromatin and RNA:DNA hybrids.

    Highlights

    Isolated from a recombinant source
    Supplied with 10X Reaction Buffer

    Product Source

    An E. Coli strain that carries an MBP fusion clone of Bovine Pancreatic DNase I.

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    DNase I Reaction Buffer-2010X

    Advantages and Features

    Applications

    • Degradation of DNA template in transcription reactions
    • Removal of contaminating genomic DNA from RNA samples
    • DNase I footprinting
    • Nick Translation

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme which will completely degrade 1 µg of pBR322 DNA in 10 minutes at 37°C in DNase I Reaction Buffer.

    Complete degradation is defined as the reduction of the majority of DNA fragments to tetranucleotides or smaller.

    Reaction Conditions

    1X DNase I Reaction Buffer
    Incubate at 37°C

    1X DNase I Reaction Buffer:
    10 mM Tris-HCl
    2.5 mM MgCl2
    0.5 mM CaCl2
    pH 7.6 @ 25°C

    Usage Concentration

    2,000 units/ml

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    2 mM CaCl2
    50% Glycerol
    pH 7.6 @ 25°C

    Heat Inactivation

    75°C for 10 min

    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.
    • RNase Activity (1 Hour Digestion):
      The product is tested in a reaction containing a RNA substrate.  After incubation for 1 hour there is no detectable degradation of the RNA substrate as determined by gel electrophoresis.

    Notes

    1. EDTA should be added to a final concentration of 5 mM to protect RNA from being degraded during enzyme inactivation (3).

    References

    1. Kunitz, M. (1950). J. Gen. Physiol . 33, 349-362.
    2. Vanecko, S. and laskowski, M. (1961). J. Biol. Chem . 236, 3312-3316.
    3. Huang, Z. et al. (1996). Biotechniques . 20, 1012-1020.

    Supporting Documents

    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]

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    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.
    1. Will DNase I work in NEB buffers 1-4?
    2. What is the best way to remove DNase I from my reaction?
    1. A Typical DNase I Reaction

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