SacI |NEB酶试剂 New England Biolabs

上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。

产品信息

本酶提供高保真版 SacI-HF®(NEB #R3156)。

高保真(HF)限制性内切酶在 rCutSmart 缓冲液中具有 100% 活性;统一缓冲液意味着更加直接、简化的样品处理过程。HF 内切酶还会显著降低星号活性。所有 HF 内切酶均符合省时酶(Time-Saver)标准,可在 5-15 分钟内酶切底物 DNA,也可实现过夜酶切,而且不会造成 DNA 降解。HF 限制性内切酶在改造时将性能作为重要指标,可在更宽的条件下具有完全活性,最大限度地减少非特异性酶切产物,并且为实验设计提供灵活性。

产品来源

大肠杆菌菌株,携带来自不产色链霉菌(Streptomyces achromogenes)(ATCC 12767)的 SacI 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,37℃ 条件下,1 小时内酶切 1 µg λ DNA(HindIII 消化)所需的酶量。

    反应条件

    1X NEBuffer™ r1.1
    Incubate at 37°C

    1X NEBuffer™ r1.1
    10 mM Bis-Tris-Propane-HCl
    10 mM MgCl2
    100 µg/ml 重组白蛋白
    (pH 7 @ 25°C)

    在不同缓冲液中的活性

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

    稀释兼容性

    • 稀释液 A

    贮存溶液

    10 mM Tris-HCl
    100 mM NaCl
    1 mM DTT
    0.1 mM EDTA
    200 µg/ml BSA
    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: 不敏感
    dcm 甲基化: 不敏感
    CpG甲基化: 不敏感

    同裂酶

    Ecl136II
    Eco53kI
    EcoICRI
    Psp124BI
    SacI-HF
    SstI

  • 相关产品

    相关产品

    • SacI-HF®
    • 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. 当盐浓度高于 10 mM 时,SacI 的活性受到抑制。小提 DNA 含残留盐分会抑制 SacI 的酶切活性。用 70% 乙醇漂洗或透析可以除去盐分。
    2. 该酶在切割某些超螺旋质粒时活性较低,酶切 1 μg 质粒 DNA 需要超过 1 个单位的酶量。如需完全酶切 1 μg 质粒 DNA,请采用我们推荐的酶切方案。
    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. Restriction Digest Protocol
    3. 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
    • 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
    • Single Letter Codes
    • Star Activity
    • Traditional Cloning Quick Guide

工具 & 资源

  • 选择指南

    • Alphabetized List of Recognition Sequences
    • Cleavage of Supercoiled DNA
    • 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. Why isn’t SacI cutting?
    2. How can the efficiency of SacI be increased?
    3. Is there any difference in the methylation sensitivity between SacI-HF and SacI?
    4. Is SacI affected by methylation?
    5. What is the molecular weight of SacI?
    6. What is the activity of SacI at 25°C?
    7. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    8. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
    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