DNase I (RNase-free) | NEB (2024)

HomeDNA Modifying Enzymes and Cloning TechnologiesProductsDNase I (RNase-free)

• Product Information

  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.

  This product is related to the following categories:

  Exonucleases and Non-specific Endonucleases Products,

  RNA Synthesis In vitro Transcription (IVT)

  • Reagents Supplied

    The following reagents are supplied with this product:

    NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
    M0303S -20 DNase I (RNase-free) M0303SVIAL -20 1 x 0.5 ml 2,000 units/ml DNase I Reaction Buffer B0303SVIAL -20 1 x 1.5 ml 10 X
    M0303L -20 DNase I (RNase-free) M0303LVIAL -20 2 x 1.25 ml 2,000 units/ml DNase I Reaction Buffer B0303SVIAL -20 1 x 1.5 ml 10 X

  • Properties & 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.

    See Also

    Working with RNA - USNucleic acid protocols: Extraction and optimization

    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 Buffer

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

    Heat Inactivation

    75°C for 10 minutes

  • Advantages and Features

    Application Features

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

  • Related Products

    Materials Sold Separately

    • DNase I Reaction Buffer

  • Product Notes

    1. EDTA should be added to a final concentration of 5 mM to protect RNA from being degraded during enzyme inactivation (3).
    2. We do not recommend using NEB's DNase I as a substitute for Monarch DNase I in the RNA isolation workflow when using the Monarch Total RNA Miniprep Kit (NEB #T2010). Monarch DNase I is optimized for use in this workflow, and is available for purchase as a component of the Monarch Total RNA Enzyme Pack, (NEB #T2019).

  • 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.

• Protocols, Manuals & Usage

  • Protocols

    1. A Typical DNase I Reaction Protocol (M0303)

  • Usage Guidelines

    • Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

• Tools & Resources

  • Selection Charts

    • Activities of Exonucleases and Non-specific Endonucleases
    • Common Applications for Exonucleases and Endonucleases
    • Properties of Exonucleases and Non-specific Endonucleases

  • Web Tools

    • Exo Selector

• FAQs & Troubleshooting

  • FAQs

    1. What is the specific activity of DNase I(RNase-free)?
    2. Will DNase I work in NEB buffers 1-4?
    3. What is the best way to remove DNase I from my reaction?
    4. Will DNase I work in rCutSmart buffer?
    5. Can I use the Monarch RNA Cleanup Kit to cleanup up my DNase I-treated RNA?
    6. Can I use NEB DNase I (#M0303) with the Monarch Total RNA Miniprep Kit?

• Citations & Technical Literature

  • Citations

    Product Citation Tool

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• Quality, Safety & Legal

  • Quality Control Assays

    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 & Change Notifications

    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]

    • M0303S_L_v1

  • 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]

    • M0303S_L_v1_0111403
    • M0303S_L_v1_0111408
    • M0303S_L_v1_0111501
    • M0303S_L_v1_0141504
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    • M0303L_v1_10226626
    • M0303S_v1_10234515
    • M0303L_v1_10234125

  • Safety Data Sheets

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

    • DNase I (RNase-free)

    • DNase I Reaction Buffer

  • Legal and Disclaimers

    Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

    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.