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
高保真(HF)限制性内切酶在 rCutSmart 缓冲液(10X)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不会降解 DNA。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,携带有克隆自粘液奈瑟菌(Neisseria mucosa heidelbergensis)(ATCC 25999)的 NheI 基因。
- 产品类别:
- Discontinued (<3 years)
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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 酶切)所需的酶量。
反应条件
1X NEBuffer™ 2.1
Incubate at 37°C1X NEBuffer™ 2.1
50 mM NaCl
10 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: 100%
NEBuffer™ 2.1: 100%
NEBuffer™ 3.1: 10%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
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: 某些甲基化与酶切位点的重叠组合阻断酶切 -
相关产品
相关产品
- NheI-HF®
- t1010-monarch-plasmid-miniprep-kit
- Monarch® DNA 胶回收试剂盒
- Monarch® PCR & DNA 纯化试剂盒(5 μg)
- Nuclease-free Water
单独销售的组分
- 6X 紫色凝胶上样染料
- NEBuffer 2.1
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注意事项
- 酶切产生的 5´ CTAG 突出末端可以与 AvrII、SpeI 或 XbaI 酶切产生的 DNA 片段有效地连接。
- 当盐浓度 > 100 mM 时,酶活性被抑制。
- 该酶在酶切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。
- 某些 CpG 甲基化与酶切位点的重叠组合阻断酶切。
操作说明、说明书 & 用法
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操作说明
- Optimizing Restriction Endonuclease Reactions
- Restriction Digest Protocol
- Double Digest Protocol with Standard Restriction Enzymes
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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
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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?
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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
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FAQs
- Is NheI affected by methylation?
- Are more units of NheI required to cut supercoiled DNA than lambda DNA?
- Is NheI inhibited by salt?
- Is NheI active at 25°C?
- Does NheI produce commonly used compatible ends?
- Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
- Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
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问题解决指南
- Restriction Enzyme Troubleshooting Guide