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造成 DNA 降解。HF 限制性内切酶在开发时便将性能作为重要指标,可在更宽的条件范围下具有完全活性,最大限度地减少非特异性酶切产物,同时为实验设计提供灵活性。
产品来源
大肠杆菌菌株,携带有克隆自球衣菌属(Sphaerotilus)(ATCC 13923)的 SpeI 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 pXba-XbaI DNA 所需的酶量。
反应条件
1X rCutSmart™ 缓冲液
Incubate at 37°C1X 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: 75%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 25%
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
pH 7.4 @ 25°C热失活
80°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 -
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
- 6X 紫色凝胶上样染料
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注意事项
- 切割产生的 5´ CTAG 突出末端能有效地和 AvrII、NheI 或 XbaI 切割生成的 DNA 片段连接。
- 对 CpG、dcm 或 dam 甲基化均不敏感。
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
- 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
- Can SpeI be used at alternative reaction temperatures?
- Is SpeI active at 25°C?
- Does SpeI 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?
- 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?
- Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
- Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
- Are there any common problems encountered when using SpeI?
- Does NEB offer any BSA-free and/or animal origin-free restriction enzymes for linearization of plasmids for mRNA vaccine development?
- Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
- Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
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问题解决指南
- Restriction Enzyme Troubleshooting Guide