BsaI |NEB酶试剂 New England Biolabs

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BsaI

This product was discontinued on 12/31/2020 and replaced with the High Fidelity version, BsaI-HFv2 (NEB #R3733).

  • Catalog # R0535 was discontinued on December 31, 2020
BsaI |
同裂酶 | 单字母标示 | Pronunciation: BsaI |

Attention Golden Gate Assembly users: BsaI-HFv2 (NEB #R3733) has been optimized for Golden Gate Assembly*.  BsaI-HFv2 also works well for any protocol requiring DNA cutting by BsaI. This is the recommended enzyme for any purpose requiring digestion at the recognition sequence …5′-GGTCTC(N1)/(N5)-3′…
 
*The requirements for digestion of DNA during Golden Gate Assembly are more demanding than what is encountered in traditional DNA cloning. For Golden Gate, the enzyme must function well in a buffer that might not be the same buffer as that normally provided for the restriction enzyme, and maintain activity for an extended time at elevated temperatures in a dynamic cutting/re-ligating reaction with competition for substrate binding between the endonuclease and ligase. Extensive testing has demonstrated superior performance of BsaI-HFv2 compared to both BsaI (NEB #R0535) and BsaI-HF (NEB  

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BsaI |

Golden Gate Assembly Workflow

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产品信息

BsaI has a High Fidelity version BsaI-HF®v2 (NEB #R3733).

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.

产品来源

An E. coli strain that carries the cloned BsaI gene from Bacillus stearothermophilus 6-55 (Z. Chen)

产品类别:
Discontinued Products

  • 特性和用法

    单位定义

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

    反应条件

    1X CutSmart® Buffer
    Incubate at 37°C

    1X CutSmart® Buffer
    50 mM Potassium Acetate
    20 mM Tris-acetate
    10 mM Magnesium Acetate
    100 µg/ml BSA
    (pH 7.9 @ 25°C)

    在不同缓冲液中的活性

    NEBuffer™ 1.1: 75%
    NEBuffer™ 2.1: 75%
    NEBuffer™ 3.1: 100%

    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

    65°C for 20 min

    甲基化敏感性

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

  • 相关产品

    相关产品

    • dam–/dcm– Competent E. coli
    • Monarch® Plasmid Miniprep Kit 
    • Monarch® DNA Gel Extraction Kit
    • Monarch® PCR & DNA Cleanup Kit (5 μg)

    单独销售的组分

    • Gel Loading Dye, Purple (6X)
    • CutSmart® Buffer

  • 注意事项
    1. Blocked by overlapping dcm methylation. Cleavage of mammalian genomic DNA is blocked by some combinations of overlapping CpG methylation.
    2. Activity at 50°C is 100%.
    3. May exhibit star activity in NEBuffer r1.1.
    4. Based on the stability of the enzyme in the reaction, incubations longerthan 1 hr will not result in improved digestion, unless additionalenzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.
    5. star activity may result from a glycerol concentration of >5%

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
    • Dam and Dcm Methylases of E. coli
    • 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
    • Technical Tips For Optimizing Golden Gate Assembly Reactions
    • Traditional Cloning Quick Guide

  • 应用实例
    • Breaking through the Limitations of Golden Gate Assembly

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
    • Enzymes with Nonpalindromic Sequences
    • Frequencies of Restriction Sites
    • Isoelectric Points (pI) for Restriction Enzymes
    • Isoschizomers
    • NEB Diluent and Buffer Table
    • Time-Saver™ Qualified Enzymes
    • Type IIS Restriction 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. Which restriction enzymes are used in Golden Gate Assembly?
    2. Which restriction enzymes are used in GoldenBraid Assembly?
    3. Why is my Restriction Enzyme not cutting DNA?
    4. Why do I see a DNA smear on an agarose gel after a restriction digest?
    5. Why do I see additional DNA bands on my gel after a restriction digest?
    6. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
    7. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
    8. What is the activity of the Type IIS restriction enzyme BsaI-HFv2 (NEB #R3733) in T4 DNA Ligase Buffer?
    9. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
    10. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?

  • 问题解决指南
    • Restriction Enzyme Troubleshooting Guide

  • 实验技巧
    Restriction Enzymes in Golden Gate Assembly
     
    BsaI |

引用 & 技术文献

  • 产品手册
    • Golden Gate Assembly brochure

  • 引用文献

    产品引用文献查找工具

    BsaI | Powered by Bioz See more details on Bioz

    更多引用文献

    • Lee JH, Won HJ, Oh E-S, Oh M-H and Jung JH (2020) Golden Gate Cloning-Compatible DNA Replicon/2A-Mediated Polycistronic Vectors for Plants Front Plant Sci; 11, 559365. DOI: 10.3389/fpls.2020.559365
    • Feng Y, Zhang S, Huang X (2014) A robust TALENs system for highly efficient mammalian genome editing Sci Rep; 4, 3632. PubMedID: 24407151, DOI: 10.1038/srep03632
    • Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T (2014) Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system Sci Rep; 4, 5400. PubMedID: 24954249, DOI: 10.1038/srep05400

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

    产品说明与变更通知

    产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]

    • R0535S_L_v2

  • 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#]

    • R0535S_L_v1_0371403
    • R0535S_L_v1_0361212
    • R0535S_L_v1_0361305
    • R0535S_L_v1_0361311
    • R0535S_L_v1_0371404
    • R0535S_L_v2_0371404
    • R0535S_L_v1_0371409
    • R0535S_L_v2_0371503
    • R0535S_L_v2_0371509
    • R0535S_L_v2_0371603
    • R0535S_L_v2_0381608
    • R0535S_L_v2_0401702
    • R0535S_L_v2_0401708
    • R0535S_L_v2_0431712
    • R0535S_L_v2_0431805
    • R0535L_v2_10009918
    • R0535S_v2_10018570
    • R0535L_v2_10030542
    • R0535S_v2_10030544
    • R0535S_v2_10043185
    • R0535L_v2_10043183
    • R0535S_v2_10055060
    • R0535S_v2_10059599
    • R0535S_v2_10062820
    • R0535L_v2_10062822
    • R0535S_v2_10065817
    • R0535S_v2_10081423
    • R0535S_v2_10091140
    • R0535L_v2_10086217

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • BsaI