NheI | NEB酶试剂 New England Biolabs

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This product has been discontinued and is replaced with the High-Fidelity version, NheI-HF (NEB #R3131)

  • Catalog # R0131 was discontinued on March 17, 2021
NheI  | NEB酶试剂 New England Biolabs
同裂酶 | 单字母标示 | Pronunciation: NheI  | NEB酶试剂 New England Biolabs
  • Time-Saver™ qualified for digestion in 5-15 minutes
  • High Fidelity (HF®) version available (NEB #R3131) supplied with CutSmart® Buffer
  • Supplied with 1 vial of Gel Loading Dye, Purple (6X)
  • Cut Site: G/CTAGC


NheI  | NEB酶试剂 New England Biolabs

Reduce Star Activity with High-Fidelity Restriction Enzymes



NheI has a High Fidelity version NheI-HF® (NEB #R3131).

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.


An E. coli strain that carries the NheI gene from Neisseria mucosa heidelbergensis (ATCC 25999).

Discontinued Products

  • 特性和用法


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


    1X NEBuffer™ 2.1
    Incubate at 37°C

    1X NEBuffer™ 2.1
    50 mM NaCl
    10 mM Tris-HCl
    10 mM MgCl2
    100 µg/ml BSA
    (pH 7.9 @ 25°C)


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


    • 稀释液 C


    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
    pH 7.4 @ 25°C


    65°C for 20 min


    dam 甲基化: Not Sensitive
    dcm 甲基化: Not Sensitive
    CpG甲基化: Blocked by Some Combinations of Overlapping

  • 相关产品


    • NheI-HF®
    • Monarch® Plasmid Miniprep Kit 
    • Monarch® DNA Gel Extraction Kit
    • Monarch® PCR & DNA Cleanup Kit (5 μg)
    • Nuclease-free Water


    • Gel Loading Dye, Purple (6X)
    • NEBuffer 2.1

  • 注意事项
    1. Cleaves to leave a 5´ CTAG extension which can be efficiently ligated to DNA fragments generated by AvrII, SpeI or XbaI.
    2. Inhibited by salt concentrations > 100 mM.
    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 some combinations of overlapping 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
    • 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
    • Site Preferences
    • 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. Is NheI affected by methylation?
    2. Are more units of NheI required to cut supercoiled DNA than lambda DNA?
    3. Is NheI inhibited by salt?
    4. Is NheI active at 25°C?
    5. Does NheI produce commonly used compatible ends?
    6. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
    7. Why is my Restriction Enzyme not cutting DNA?
    8. Why do I see a DNA smear on an agarose gel after a restriction digest?
    9. Why do I see additional DNA bands on my gel after a restriction digest?
    10. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
    11. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?

  • 问题解决指南
    • Restriction Enzyme Troubleshooting Guide

引用 & 技术文献

  • 引用文献


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质控、安全 & 法规

  • 质控分析
    每一新批次的 NEB 产品都要经过质控,以满足指定的质量标准,对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息,可从此处查阅。

  • 产品说明与变更通知



    • R0131M_v1
    • R0131S_L_v1

  • CoA
    CoA 文件包含单个批次产品的储存温度、有效期和质控,这些文件的命名规则如下: [货号]_[规格]_[版本]_[Lot#]

    • R0131M_v1_0281306
    • R0131M_v1_0291312
    • R0131S_L_v1_0281301
    • R0131S_L_v1_0291306
    • R0131S_L_v1_0291312
    • R0131M_v1_0271301
    • R0131M_v1_0291405
    • R0131S_L_v1_0271301
    • R0131S_L_v1_0291405
    • R0131M_v1_0291410
    • R0131S_L_v1_0291410
    • R0131S_L_v1_0291504
    • R0131M_v1_0291504
    • R0131S_L_v1_0291509
    • R0131S_L_v1_0291604
    • R0131M_v1_0291509
    • R0131M_v1_0291605
    • R0131S_L_v1_0291608
    • R0131S_L_v1_0291611
    • R0131S_L_v1_0291701
    • R0131M_v1_0291705
    • R0131S_L_v1_0291704
    • R0131M_v1_0291711
    • R0131S_L_v1_0291711
    • R0131M_v1_0291701
    • R0131M_v1_10011628
    • R0131L_v1_10008860
    • R0131S_v1_10015942
    • R0131S_v1_10020681
    • R0131S_v1_10027617
    • R0131M_v1_10032186
    • R0131S_v1_10035482
    • R0131L_v1_10027620
    • R0131L_v1_10042389
    • R0131S_v1_10042391
    • R0131M_v1_10053501
    • R0131S_v1_10057181
    • R0131L_v1_10063024
    • R0131S_v1_10063025
    • R0131M_v1_10071487
    • R0131S_v1_10069390
    • R0131S_v1_10078814
    • R0131S_v1_10085530
    • R0131S_v1_10091136
    • R0131L_v1_10088050
    • R0131M_v1_10088048
    • R0131S_v1_10091579

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • NheI