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)
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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 消化）所需的酶量。
反应条件
1X NEBuffer™ r1.1
Incubate at 37°C1X 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）
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注意事项
- 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。
- 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。
- 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
- 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
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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
- 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
- Why isn’t SacI cutting?
- How can the efficiency of SacI be increased?
- Is there any difference in the methylation sensitivity between SacI-HF and SacI?
- Is SacI affected by methylation?
- What is the molecular weight of SacI?
- What is the activity of SacI at 25°C?
- 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)?
- Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
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问题解决指南
- Restriction Enzyme Troubleshooting Guide
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