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Bst DNA Polymerase, Large fragment

DNA polymerase I

价格 ￥240.00 （ 供应商现货 ： 3-5个工作日）
货号 UA070065
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产品介绍

产品介绍 评论(0)

产品规格

  • 分子别名

    DNA polymerase I

  • 表达宿主

    E.coli

  • 分子量

    67kDa (Reducing)

  • 纯度

    ＞95% by SDS-PAGE

  • 活性

    8U/μl

  • 标签蛋白&酶切位点

    /

  • 标签

    His Tag

  • 缓冲体系

    50 mM KCl、10 mM Tris-HCl、0.1 mM EDTA、1 mM DTT、0.1% Triton® X-100、50% Glycerol、pH 7.1 @ 25°C

  • 储存条件

    Store at -25 ~ -15℃ for 2 years

  • 文献引用

    1. Nucleic Acids Research, 2000, 28(12):E63.
    2. Chemical Communications, 2014, 50(28):3747-3749.
    3. Current Protocols in Molecular Biology, 2014(Suppl.105):15.14.1-15.14.14.

背景介绍

  • Bst DNA Polymerase, Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the
    5´ → 3´ polymerase activity, but lacks 5´ →3´ exonuclease activity. It can be use to isothermal amplification (LAMP), DNA sequencing through high GC regions and Rapid Sequencing from nanogram amounts of DNA template.

产品组分

  • 50 mM KCl、10 mM Tris-HCl、0.1 mM EDTA、1 mM DTT、0.1% Triton® X-100、50% Glycerol、pH 7.1 @ 25°C
    10* Reaction Buffer: 200 mM Tris-HCl、100 mM (NH4)2SO4、20mM MgSO4、100 mM KCl、1% Tween® 20、pH 8.8@25°C
    Magnesium Sulfate (MgSO4) Solution：100mM MgSO4

操作步骤

  • Incubate the following reaction at 65°C for 30–60 minutes

    Component

    Final Concentration

    10X Isothermal Amplification Buffer II

    1X (contains 2 mM MgSO4)

    MgSO4 (100 mM)

    6 mM (8 mM total)

    dNTP Mix (10 mM)

    1.4 mM each

    FIP/BIP Primers (25X)

    1.6 µM

    F3/B3 Primers (25X)

    0.2 µM

    LoopF/B Primers (25X)

    0.4 µM

    Bst DNA Polymerase (8,000 U/ml)

    320 U/ml

    DNA or RNA Sample

    > 10 copies or more

    Nuclease-free Water

    to 25 µl

    Total Reaction Volume

    25 µl



注意事项

  • 1.Bst DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.

    2.100 µg/ml BSA or 0.1%Triton X-100 is required for long term storage.

    3.Reaction temperatures above 70°C are not recommended.

    4.Bst DNA Polymerase, Large Fragment cannot be used for thermal cycle sequencing or PCR

酶活定义
  • One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 65°C

  • 生物活性

    • Experimental design, in a 25μl system, using the same amount (0.1ng) of pCMV-Cre-EGFP as the template, using different amounts of this product or Bst DNA polymerase of Competitor, primer: 1.6µM FIP/BIP, 0.2µM F3/B3, 0.4µM Loop F/B, 1.4mM dNTP each, MgSO4 was added to 1X Bst Reaction Buffer (2mM MgSO4) until the final concentration reached 8mM, and incubated at 65ºC for 1 hour. Inactivation was heated at 80ºC for 20 minutes and then tested by 2.0% agarose gel electrophoresis.
      As shown in the figure, this product has comparable enzyme activity compared with Competitor N's products.
      Lane 1 Negative Control-1(negative control with no added enzyme only)
      Lane 2 Negative Control-2(only negative controls without templates)
      Lane 3 UA070061- Bst DNA Polymerase, Large fragment 8U
      Lane 4 competing product N 8U

  • 电泳

    • 2μg (R: reducing condition, N: non-reducing condition).

  • 体积排阻色谱(SEC-HPLC)

    • 95.5%

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