DNase I (RNase-free) |NEB酶试剂 New England Biolabs

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产品信息

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.

重点

Isolated from a recombinant source
Supplied with 10X Reaction Buffer

产品来源

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

产品类别:
Exonucleases and Non-specific Endonucleases Products,
RNA Synthesis In vitro Transcription (IVT)

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • 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

  • 特性和用法

    单位定义

    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.

    反应条件

    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)

    使用浓度

    2,000 units/ml

    贮存溶液

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

    热失活

    75°C for 10 minutes

  • 优势和特性

    应用特性

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

  • 相关产品

    单独销售的组分

    • DNase I 反应缓冲液

  • 注意事项

    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).

  • 参考文献

    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.

操作说明、说明书 & 用法

  • 操作说明

    1. A Typical DNase I Reaction Protocol (M0303)

  • 使用指南

    • Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

工具 & 资源

  • 选择指南

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

  • Web 工具

    • Exo Selector

FAQs & 问题解决指南

  • 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?