CviAII |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，携带有来自小球藻病毒（Chlorella virus）PBCV-1（J.L. Van Etten）的 CviAII 基因。

产品类别:
Discontinued (<3 years)

  • 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，25℃ 下，1 小时内酶切 1 µg pUC19 DNA 所需的酶量。

    反应条件

    1X rCutSmart™ 缓冲液
    Incubate at 25°C

    1X rCutSmart™ 缓冲液
    50 mM Potassium Acetate
    20 mM Tris-acetate
    10 mM Magnesium Acetate
    100 µg/ml Recombinant Albumin
    (pH 7.9 @ 25°C)

    在不同缓冲液中的活性

    NEBuffer™ r1.1: 50%
    NEBuffer™ r2.1: 50%
    NEBuffer™ r3.1: 10%
    rCutSmart™ Buffer: 100%

    稀释兼容性

    • 稀释液 C

    贮存溶液

    10 mM Tris-HCl
    250 mM NaCl
    1 mM DTT
    0.1 mM EDTA
    200 µg/ml BSA
    50% Glycerol
    0.15% Triton® X-100
    10 mM TCEP
    pH 7.4 @ 25°C

    热失活

    65°C for 20 minutes

    甲基化敏感性

    dam 甲基化: 不敏感
    dcm 甲基化: 不敏感
    CpG甲基化: 不敏感

    同裂酶

    FaeI
    FatI
    Hin1II
    Hsp92II
    NlaIII

    在不同温度下的活性

    @37°C: 10%

  • 相关产品

    相关产品

    • t1010-monarch-plasmid-miniprep-kit
    • Monarch® DNA 胶回收试剂盒
    • Monarch® PCR & DNA 纯化试剂盒（5 μg）

    单独销售的组分

    • rCutSmart™ 缓冲液

  • 注意事项

    1. CviAII 是 NlaIII 的异裂酶。
    2. 因为反应中酶的稳定性，即使温育时间超过 1 小时也不会提高酶切效率，除非增加酶量。如需了解更多信息，请参阅限制性内切酶在反应中的活性。
    3. 对 CpG、dcm 或 dam 甲基化均不敏感。

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
    • Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
    • Double Digests
    • 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

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
    • 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. Is CviAII activity sensitive to dam, dcm or mammalian CpG methylation?
    2. How many base pairs should be added at the end of a PCR primer after the CviAII recognition site to guarantee that CviAII will cut properly?
    3. Does CviAII have any neoschizomers?
    4. My CviAII enzyme was working fine several months ago, but now it is not working anymore. Is there a reason?
    5. I have run my DNA digested with CviAII in a gel and I cannot see the digested fragment that I was expecting. Has the enzyme degraded my substrate DNA?
    6. What is the activity of CviAII at 37°C?
    7. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    8. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
    9. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

  • 问题解决指南

    • Restriction Enzyme Troubleshooting Guide