EagI |NEB酶试剂 New England Biolabs

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产品信息

EagI has a High Fidelity version, EagI-HF® (NEB #R3505)

High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 minutes with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design.

产品来源

An E. coli strain that carries the EagI gene from Enterobacter agglomerans (R. Morgan).

产品类别:
Discontinued (<3 years)

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to digest 1 µg of pXba DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

    反应条件

    1X NEBuffer™ 3.1
    Incubate at 37°C

    1X NEBuffer™ 3.1
    100 mM NaCl
    50 mM Tris-HCl
    10 mM MgCl2
    100 µg/ml BSA
    (pH 7.9 @ 25°C)

    在不同缓冲液中的活性

    NEBuffer™ 1.1: 10%
    NEBuffer™ 2.1: 25%
    NEBuffer™ 3.1: 100%

    稀释兼容性

    • 稀释液 B

    贮存溶液

    10 mM Tris-HCl
    500 mM NaCl
    1 mM DTT
    0.1 mM EDTA
    200 µg/ml BSA
    50% Glycerol
    pH 8 @ 25°C

    热失活

    65°C for 20 minutes

    甲基化敏感性

    dam 甲基化: Not Sensitive
    dcm 甲基化: Not Sensitive
    CpG甲基化: Blocked

    同裂酶

    XmaIII+

  • 相关产品

    相关产品

    • EagI-HF®
    • 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

    • 6X 紫色凝胶上样染料
    • NEBuffer 3.1

  • 注意事项

    1. EagI is an isoschizomer of XmaIII.
    2. For full EagI activity, the pH of the reaction mix must be between (7.9 and 9.0 @ 25°C).
    3. This enzyme has shown to have lower activity on some supercoiled plasmids, with more than 1 unit required to digest 1 μg plasmid DNA. For complete digestion of 1 μg of plasmid DNA please follow our recommended digestion protocol.
    4. Blocked by CpG methylation.

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
    • 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
    • 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
    • Recleavable Filled-in 5′ Overhangs
    • 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. Does EagI produce commonly used compatible ends?
    2. Are more units of EagI required to cut supercoiled DNA than lambda DNA?
    3. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    4. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
    5. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?

  • 问题解决指南

    • Restriction Enzyme Troubleshooting Guide