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产品中心 分子酶 mRNA疫苗

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DNase Ⅰ

DNASE,Deoxyribonuclease-1

价格 ￥455.00 （ 供应商现货 ： 3-5个工作日）
货号 UA070036
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产品介绍

产品介绍 评论(0)

产品规格

  • 物种

    Bovine Pancreatic

  • 分子别名

    DNASE,Deoxyribonuclease-1

  • 表达宿主

    E.coli

  • 分子量

    72kDa (Reducing)

  • 纯度

    >95% by SDS-PAGE&HPLC

  • 活性

    2U/ul

  • 标记

    Unconjugated

  • 标签

    MBP Tag, His Tag

  • 性状

    Liquid

  • 缓冲体系

    10 mM Tris-HCl, 2 mM CaCl2 ,50% Glycerol，(pH 7.6, 25°C)

  • 储存条件

    Store at -25 ~ -15℃ for 2 years

  • 文献引用

    [1] Vanecko S, Laskowski M. Studies of the Specificity of Deoxyribonuclease I[J]. Journal of Biological Chemistry, 1961, 236(236):3312-6.

    [2] Kienzle N, Young D, Zehntner S, et al. DNaseI treatment is a prerequisite for the amplification of cDNA from episomal-based genes[J]. Biotechniques, 1996, 20(4):612-6.

    [3] Michael,R, Green, et al. Human β-globin pre-mRNA synthesized in vitro is accurately spliced in xenopus oocyte nuclei[J].Cell, 1983, 32(3):681-694.

背景介绍

  • DNase I (Deoxyribonuclease I), can digest single or double-stranded DNA to produce mono deoxynucleotides or single or double-stranded oligo deoxynucleotides, its optimal working pH range is 7-8. DNase I activity is dependent on Ca2+ and can be activated by other bivalent metal ions such as Mg2+, Mn2+, Zn2+, etc. In the presence of Mg2+, the enzyme can randomly recognize and cut any site on any strand of DNA. In the presence of Mn2+, two strands of DNA can be cut at the same site to form sticky ends with flat ends or 1-2 nucleotides protruding.

产品组分

  • Storage Solution: 2 U/ul DnaseⅠ、10mM Tris-Hcl、2mM CaCl2、50%Glycerol (pH7.6, 25℃)

    10*Reaction Buffer: 100mM Tris-Hcl、25mM MgCl2、5mM CaCl2 (pH7.6, 25℃)

操作步骤

  • This step is suitable for linearization of 1 μg DNA (≥100 nt) and can be scaled up according to experimental needs.

    1）Add the following components in sequence

    Components

    Volume

    Plasmid DNA

    1μg DNA

    10*Reaction Buffer

    2μl

    DnaseⅠ (2U/μl)

    1μl

    RNase-free ddH2O

    Up to 50μl

    2）Incubate at 37°C 1 h.

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 2. Please avoid repeated freeze-thaw cycles

酶活定义
  • 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.

  • 生物活性

    • The results of 1μg pBR322 plasmid digestion separated under different quantity of DnaseⅠ, The reaction was incubated for 10 minutes at 37°C, and 1% agarose gel was used for electrophoresis analysis after reaction.

      M, marker;

      Lane 1 1μg pBR322;

      Lane 2 1μg pBR322 add 4U DNase I

      Lane 3 1μg pBR322 add 2U DNase I

      Lane 4 1μg pBR322 add 1U DNase I

      Lane 5 1μg pBR322 add 0.5U DNase I

      Lane 6 1μg pBR322 add 0.25U DNase I

      Lane 7 1μg pBR322 add 0.125U DNase I

  • 电泳

    • 1μg (R: reducing condition, N: non-reducing condition).

  • 反相高效液相色谱(RP-HPLC)

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