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,携带有克隆自普通变形杆菌(Proteus vulgaris)(ATCC 13315)的 PvuI 基因
- 产品类别:
- Discontinued (<3 years)
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产品组分信息
本产品提供以下试剂或组分:
NEB # 名称 组分货号 储存温度 数量 浓度 -
R0150V -20 PvuI R0150VVIAL -20 1 x 0.025 ml 10,000 units/ml NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X
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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 DNA 所需的酶量。
反应条件
1X NEBuffer™ r3.1
Incubate at 37°C1X NEBuffer™ r3.1
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)在不同缓冲液中的活性
NEBuffer™ r1.1: <10%
NEBuffer™ r2.1: 25%
NEBuffer™ r3.1: 100%
rCutSmart™ Buffer: <10%稀释兼容性
- 稀释液 B
贮存溶液
10 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
500 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C热失活
否
甲基化敏感性
dam 甲基化: 不敏感
dcm 甲基化: 不敏感
CpG甲基化: 阻断酶切 -
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)
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注意事项
- 哺乳动物基因组 DNA CpG 甲基化阻断酶切。
- 对于不能热失活的酶,我们建议进行柱纯化(如 Monarch® PCR & DNA 纯化试剂盒),或琼脂糖凝胶电泳后切胶回收(建议使用 Monarch DNA 胶回收试剂盒),或酚/氯仿抽提。
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
- 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
- Is PvuI activity sensitive to dam, dcm or mammalian CpG methylation?
- How many base pairs should be added at the end of a PCR primer after the PvuI recognition site to guarantee that PvuI will cut properly?
- Does PvuI have any neoschizomers?
- 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