NEB酶试剂 New England Biolabs® Turbo Electrocompetent E. coli | NEB酶试剂 New England Biolabs

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NEB® Turbo Electrocompetent E. coli

E. coli cells featuring fast colony growth (6.5 hours)

  • Tight expression control (lacIq)
  • Isolate DNA after only 4 hours of growth
  • 5-minute transformation protocol with AmpR plasmids
  • No dry ice surcharge on competent cell shipments
  • Outgrowth medium included
  • Free of animal products
  • Refer to our Electroporation Tips

精选视频

NEB酶试剂 New England Biolabs® Turbo Electrocompetent E. coli | NEB酶试剂 New England Biolabs

Differences between electrocompetent and chemically competent cells

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产品信息

NEB Turbo Electrocompetent E. coli cells are suitable for high efficiency electroporation and rapid colony growth. These cells are ideal for DNA library constructions and all cloning purposes.

重点

  • Transformation efficiency >1 x 1010 cfu/μg pUC19 DNA
  • Tight control of expression by lacIq allows potentially toxic genes to be cloned
  • Highest growth rate on agar plates – visible colonies 6.5 hours after transformation
  • Activity of nonspecific endonuclease I (endA) eliminated for highest quality plasmid preparations
  • Resistance to phage T1 (fhuA2)
  • Suitable for blue/white screening by α-complementation of the β-galactosidase gene
  • Isolate DNA after 4 hours of growth from a single overnight colony
  • EcoKr–m–, McrBC–
  • K12 Strain

基因型

F’ proA+B+ lacIq ∆lacZM15 / fhuA2  ∆(lac-proAB)  glnV galK16 galE15  R(zgb-210::Tn10)TetS  endA1 thi-1 ∆(hsdS-mcrB)5  

产品类别:
Discontinued Products

  • 特性和用法

    Antibiotic for Plasmid Selection

    用于质粒筛选的抗生素 使用浓度
    Ampicillin 100 µg/ml
    Carbenicillin 100 µg/ml
    Chloramphenicol 33 µg/ml
    Kanamycin 30 µg/ml
    Streptomycin 25 µg/ml
    Tetracycline 15 µg/ml

    货运单

    • Ships on dry ice

    抗生素耐药性

    • nit

  • 优势和特性

    Features

    Electroporation Tips

    1. Electroporation cuvettes and microcentrifuge tubes should be pre-chilled on ice. 
    2. Electrocompetent cells should be thawed on ice and suspended well by carefully flicking the tubes. 
    3. Once DNA is added to the cells, electroporation can be carried out immediately. It is not necessary to incubate DNA with cells. The maximum recommended volume of a DNA solution to be added is 2.5 µl. Addition of a large volume of DNA decreases transformation efficiency. 
    4. Contaminants such as salts and proteins can lower electroporation efficiency. Ideally, DNA for transformation should be purified and suspended in water or TE. Transformation efficiency is more than10-fold lower for ligation mixtures than the control pUC19 plasmid due to the presence of ligase and salts. If used directly, ligation reactions should be heat-inactivated at 65°C for 20 min and then diluted 10-fold. For optimal results, spin columns (NEB #T1030) are recommended for cleanup of ligation reactions. 
    5. Electroporation conditions vary with different cuvettes and electroporators. If you are using electroporators or cuvettes not specified in the protocol, you may need to optimize the electroporation conditions. Cuvettes with 1mm gap are recommended (e.g. BTX Model 610/613 and Bio-Rad #165-2089). Higher voltage is required for cuvettes with 2 mm gap. 
    6. Arcing may occur due to high concentration of salts or air bubbles. 
    7. It is essential to add recovery medium to the cells immediately after electroporation. One minute delay can cause a 3-fold reduction in efficiency. 
    8. Cold and dry selection plates lead to lower transformation efficiency. Pre-warm plates at 37°C for 1 hour. Using 37°C pre-warmed recovery medium increases the efficiency by about 20%. 
    9. Refreeze unused cells in a dry ice/ethanol bath for 5 min and then store at -80°C. Do not use liquid nitrogen. Additional freeze-thaw cycles result in lower transformation efficiency.

    应用特性

    DNA Effects of Transformation Efficiency and Colony Output:NEB酶试剂 New England Biolabs® Turbo Electrocompetent E. coli | NEB酶试剂 New England Biolabs
    Electroporation efficiency remains extremely high up to about 1 ng of input DNA, then decreases at higher DNA concentrations.

  • 相关产品

    相关产品

    • NEB® 10-beta Electrocompetent E. coli
    • NEB® Turbo Competent E. coli (High Efficiency)
    • SOC Outgrowth Medium
    • ElectroLigase®
    • phi29 DNA Polymerase
    • Monarch® Plasmid Miniprep Kit 
    • Monarch® DNA Gel Extraction Kit
    • Monarch® PCR & DNA Cleanup Kit (5 μg)

  • 注意事项
    1. STORAGE AND HANDLING: Competent cells should be stored at -80°C. Storage at -20°C will result in a significant decrease in transformation efficiency. Cells lose efficiency whenever they are warmed above -80°C, even if they do not thaw.

操作说明、说明书 & 用法

  • 操作说明
    1. Electroporation Protocol (C2986)

  • 使用指南
    • Additional E. coli Strain Genotypes
    • Electroporation Tips
    • Genetic Markers
    • McrA, McrBC and EcoKI Strain Phenotypes
    • Traditional Cloning Quick Guide

工具 & 资源

  • 选择指南
    • Characteristics of Select E.coli Strains

  • Web 工具
    • Competitor Cross-Reference Tool
    • NEBcloner®

FAQs & 问题解决指南

  • FAQs
    1. How should I calculate the Electrotransformation efficiency (C2986)?
    2. What are the solutions/recipes (C2986)?
    3. What are the strain properties (C2986)?
    4. Can any NEB competent cells help with the generation of custom phage libraries? An F factor is required for such libraries.
    5. How should I store SOC Outgrowth Medium? The SOC I received with my competent cells recommends storage at either room temperature or 4°C, however, when I purchase it as a stand alone product, it recommends storing it at 4°C. Which is better?
    6. How should fragments be prepared for assembly using NEBuilder HiFi?
    7. Are NEB Turbo Competent E. coli (High Efficiency) or NEB Turbo Electrocompetent E. coli recommended for cloning repetitive or unstable DNA?

引用 & 技术文献

  • 引用文献

    产品引用文献查找工具

    NEB酶试剂 New England Biolabs® Turbo Electrocompetent E. coli | NEB酶试剂 New England Biolabs Powered by Bioz See more details on Bioz

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]

    • C2986K_v1

  • 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#]

    • C2986K_v1_0221612
    • C2986K_v1_0221703
    • C2986K_v1_0231704
    • C2986K_v1_0231707
    • C2986K_v1_0231711
    • C2986K_v1_0231712
    • C2986K_v1_0231801
    • C2986K_v1_0241802
    • C2986K_v1_0241803
    • C2986K_v1_10013427
    • C2986K_v1_10022955
    • C2986K_v1_10025667
    • C2986K_v1_10029521
    • C2986K_v1_10031962
    • C2986K_v1_10032691
    • C2986K_v1_10034657
    • C2986K_v1_10036358
    • C2986K_v1_10042671
    • C2986K_v1_10047055
    • C2986K_v1_10049341
    • C2986K_v1_10054346
    • C2986K_v1_10055903
    • C2986K_v1_10058816
    • C2986K_v1_10060354
    • C2986K_v1_10063869
    • C2986K_v1_10069372
    • C2986K_v1_10070200
    • C2986K_v1_10074555
    • C2986K_v1_10074997
    • C2986K_v1_10077690
    • C2986K_v1_10081196
    • C2986K_v1_10083933
    • C2986K_v1_10086890
    • C2986K_v1_10091481

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • NEB® Turbo Electrocompetent E. coli