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产品中心 分子酶 分子诊断&NGS测序相关

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Ultra Nuclease (Tag free) 全能核酸酶

Nuclease,Ultra Nuclease,Benzonase、Endonuclease

价格 ￥420.00 （ 供应商现货 ： 3-5个工作日）
货号 UA070057
规格
数量
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产品介绍 评论(0)

产品规格

  • 物种

    Serratia marcescens

  • 分子别名

    Nuclease, Ultra Nuclease, Benzonase、Endonuclease

  • 表达宿主

    E.coli

  • 分子量

    27kDa (Reducing)

  • 纯度

    ＞95% by SDS-PAGE and HPLC

  • 活性

    ≥250U/μl

  • 标记

    Unconjugated

  • 性状

    Liquid

  • 缓冲体系

    10mM Tris (pH7.4), 500mM NaCl, 2mM MgCl2, 50% glycerol

  • 溶解方法

    /

  • 储存条件

    Store at -25 ~ -15℃ for 2 years

  • 文献引用

    1. Nestle M, Roberts W K. An Extracellular Nuclease from Serratia marcescens I. PURIFICATION AND SOME PROPERTIES OF THE ENZYME[J]. Journal of Biological Chemistry, 1969, 244.
    2. Kim W Y, Lee H S, Suh S J, et al. Purification and Cellular Localization of Extracellular Nuclease of Serratia marcescens Expressed in Escherichia coli[J]. Korean Journal of Microbiology, 1994, 32(2):147-154.

  • 稀释度

背景介绍

  • Multi Nuclease also called broad-spectrum nucleic acid enzyme, is a kind of comes from Serratia Marcescens restriction endonuclease. It is capable of degradation of all forms of DNA and RNA (double-stranded, single-stranded, linear, circular or super helical forms) under a very wide range of conditions (6Murea, 0.1M Guanidine, 0.4%TritonX100, 0.1%SDS, 1mM EDTA, 1mM PMSF). The expression and purification of this product in Escherichia coli(E.coli) through genetic engineering can not only reduce the viscosity of cell supernatant and cell lysate in scientific research, but also improve the efficiency of protein purification and functional research. It can also be used in virus purification, vaccine production, protein and polysaccharide pharmaceutical industry as a host residual nucleic acid removal reagent, reducing the host residual nucleic acid to the peak (pg) level to improve the efficacy and safety of biological products. And can effectively prevent human peripheral blood monocyte (PBMC) clumping in cell therapy and vaccine research.

产品组分

  • Storage Solution: ≥250U/μl Ultra Nuclease (Tag free)、10mM Tris (pH7.4), 500mM NaCl, 2mM MgCl2, 50% glycerol

操作步骤

  • 1. Sample preparation:

    Adherent cells: Remove the medium, clean the cells with PBS, and remove the supernatant.

    Suspension cells: Cells were collected by centrifugation, cleaned with PBS, centrifuged at 6,000rpm for 10min, and precipitates were collected.

    Escherichia coli: The bacteria were collected by centrifugation, cleaned once with PBS, centrifuged at 8,000rpm for 5min, and precipitates were collected.

    2. Sample treatment:

    The collected cell precipitates are cleaved according to the ratio of mass (g) to volume (mL) to 1: (10~20). Cells can also be cleaved mechanically or chemically on ice or at room temperature (1g cells are about 109).

    3. Enzyme addition:

    the proportion of 1g cell precipitation digested by 250Units is required. You can also choose the addition plan according to the recommended dosage in the table above, increase the amount of enzyme within a certain range, and reduce the digestion time accordingly.

    4. Supernatant acquisition:

    The supernatant of cell lysis solution was obtained by centrifugation at 12,000rpm for 30min, and then subsequent related experiments were conducted.

注意事项

  • Recommended reaction condition:

    Conditional parameter

    Optimum condition

    Applicable condition

    Mg2+

    1-2mM

    1-10mM

    PH

    8.0

    6-10

    Temperature

    37℃

    0-42℃

    DTT

    0-100mM

    >0mM

    β-Me

    0-100mM

    ＞0mM

    Monovalent cation

    0-20mM

    0-150mM

    phosphate anion

    0-10mM

    0-100mM


酶活定义
  • One unit of Nuclease is defined as the amount of enzyme that causes a ∆A260 of 1.0 (equivalent to the complete digestion of 37μg DNA) in 30min

  • 生物活性

    • M marker

      1: Expi 293 Cell Lysis solution A(ultrasonicated).

      2: Expi 293 Cell Lysis solution B( non-ultrasonicated).

      3: Expi 293 Cell Lysis solution A+0.5μl Ultra Nuclease (Tag free).

      4: Expi 293 Cell Lysis solution B+0.5μl Ultra Nuclease(Tag free).

      5: Expi 293 Cell Lysis solution A+1μl Ultra Nuclease(Tag free).

      6: Expi 293 Cell Lysis solution B+1μl Ultra Nuclease(Tag free).

      Reaction condition: Incubate at 37℃ for 30mins.

    • M marker

      1: plasmid A.

      2:plasmid A+ Ultra Nuclease(Tag free).

      3:plasmid B.

      4:plasmid B+ Ultra Nuclease(Tag free).

  • 电泳

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

  • 反相高效液相色谱(RP-HPLC)

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