DpnII |NEB酶试剂 New England Biolabs

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产品信息

产品来源

大肠杆菌菌株，携带来自肺炎双球菌（Diplococcus pneumoniae）G41（S. Lacks）的 DpnII 基因。

产品类别:
Methylation Sensitive Restriction Enzymes for Epigenetics Products,
Restriction Enzymes for Epigenetic Analysis,
Restriction Endonucleases C G Products,

Epigenetic Analysis Products,

Time-Saver Qualified Restriction Enzymes Products

应用:
Restriction Enzymes for Epigenetics,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB,
Restriction Enzyme Digestion

  • 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
    • R0543S     -20    
        DpnII R0543SVIAL -20 1 x 0.1 ml 10,000 units/ml
        NEBuffer™ DpnII B0543SVIAL -20 1 x 1.25 ml 10 X
        Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
    • R0543T     -20    
        DpnII R0543TVIAL -20 1 x 0.02 ml 50,000 units/ml
        NEBuffer™ DpnII B0543SVIAL -20 1 x 1.25 ml 10 X
        Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
    • R0543L     -20    
        DpnII R0543LVIAL -20 1 x 0.5 ml 10,000 units/ml
        NEBuffer™ DpnII B0543SVIAL -20 1 x 1.25 ml 10 X
        Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
    • R0543M     -20    
        DpnII R0543MVIAL -20 1 x 0.1 ml 50,000 units/ml
        NEBuffer™ DpnII B0543SVIAL -20 1 x 1.25 ml 10 X
        Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X

  • 特性和用法

    单位定义

    一个单位是指在 50 µl 的总反应体系中，37℃ 下，1 小时内酶切 1 µg λ DNA（dam-）所需的酶量。

    反应条件

    1X NEBuffer™ DpnII
    Incubate at 37°C

    1X NEBuffer™ DpnII
    50 mM Bis-Tris-HCl
    100 mM NaCl
    10 mM MgCl2
    0.1 mM DTT
    (pH 6 @ 25°C)

    在不同缓冲液中的活性

    NEBuffer™ r1.1: 25%
    NEBuffer™ r2.1: 25%
    NEBuffer™ r3.1: 100%
    rCutSmart™ Buffer: 25%

    稀释兼容性

    • 稀释液 B

    贮存溶液

    10 mM Tris-HCl
    300 mM NaCl
    1 mM DTT
    0.1 mM EDTA
    500 µg/ml Recombinant Albumin
    50% Glycerol
    pH 7.4 @ 25°C

    热失活

    65°C for 20 minutes

    甲基化敏感性

    dam 甲基化: 阻断酶切
    dcm 甲基化: 不敏感
    CpG甲基化: 不敏感

    同裂酶

    Bsp143I
    BssMI
    BstKTI
    BstMBI
    Kzo9I
    MboI
    NdeII
    Sau3AI

  • 相关产品

    相关产品

    • dam–/dcm– E. coli 感受态细胞
    • t1010-monarch-plasmid-miniprep-kit
    • Monarch® DNA 胶回收试剂盒
    • Monarch® PCR & DNA 纯化试剂盒（5 μg）

  • 注意事项

    1. DpnII 和 Sau3AI 是 MboI 的完全同裂酶。
    2. 切割产生 5´ GATC 突出末端，该末端可高效地连接由 BamHI、BclI、BglII、MboI、Sau3AI 和 BstYI 酶切生成的 DNA 片段。
    3. dam 甲基化阻断该酶酶切。如需了解更多信息，请参阅 Dam-Dcm 和 CpG 甲基化。
    4. 在 NEBuffer r3.1 缓冲液中可能出现星号活性。推荐使用 NEB DpnII 专用缓冲液。

操作说明、说明书 & 用法

  • 操作说明

    1. Optimizing Restriction Endonuclease Reactions
    2. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
    3. Restriction Digest Protocol
    4. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
    5. Double Digest Protocol with Standard Restriction Enzymes

  • 使用指南

    • Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
    • Activity of Restriction Enzymes in PCR Buffers
    • Cleavage Close to the End of DNA Fragments
    • Dam and Dcm Methylases of E. coli
    • Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
    • Double Digests
    • Effects of CpG Methylation on Restriction Enzyme Cleavage
    • Heat Inactivation
    • NEBuffer Activity/Performance Chart with Restriction Enzymes
    • Optimizing Restriction Endonuclease Reactions
    • Restriction Endonucleases – Survival in a Reaction
    • Restriction Enzyme Diluent Buffer Compatibility
    • Restriction Enzyme Tips
    • Restriction Enzymes for Droplet Digital PCR (ddPCR)
    • Single Letter Codes
    • Star Activity
    • Traditional Cloning Quick Guide

工具 & 资源

  • 选择指南

    • Alphabetized List of Recognition Sequences
    • Compatible Cohesive Ends and Generation of New Restriction Sites
    • Dam-Dcm and CpG Methylation
    • Frequencies of Restriction Sites
    • Isoelectric Points (pI) for Restriction Enzymes
    • Isoschizomers
    • NEB Diluent and Buffer Table
    • Recleavable Filled-in 5′ Overhangs
    • Restriction Enzymes for Epigenetics Selection Chart
    • Time-Saver™ Qualified Enzymes
    • Why Choose Recombinant Enzymes?

  • Web 工具

    • Competitor Cross-Reference Tool
    • DNA Sequences and Maps Tool
    • Double Digest Finder
    • Enzyme Finder
    • NEBcutter™ v3.0
    • NEBioCalculator®
    • REBASE®

FAQs & 问题解决指南

  • FAQs

    1. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    2. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
    3. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
    4. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?

  • 问题解决指南

    • Restriction Enzyme Troubleshooting Guide

  • 实验技巧

    此酶的切割会被 dam 甲基化位点阻断，因此不能酶切从大多数大肠杆菌中提取的质粒。只有从大肠杆菌的 dam-、dcm- 菌株中转化并提取的质粒方可被此酶切割，例如 NEB 的 dam-/dcm- 大肠杆菌感受态细胞（NEB #C2925）。