10-beta E. coli 感受态细胞(高效级) |NEB酶试剂 New England Biolabs

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产品信息

E. coli 化学感受态细胞适用于多种应用中的高效转化。

重点

  • DH10B™ 衍生菌株
  • 可高效转化真核生物来源的甲基化 DNA,或来自 PCR、cDNA 和其它来源 [mcrAΔ(mrr-hsdRMS-mcrBC)] 的非甲基化 DNA
  • 去除了非特异性核酸内切酶 I(endA1)的活性,以获得最高质量的质粒制备
  • 抗 T1 噬菌体(fhuA)
  • 适用于 b-半乳糖苷酶基因的 a-互补蓝白斑筛选(无 IPTG)
  • 减少克隆 DNA 的重组(recA1
  • 可克隆大质粒和 BAC
  • 无动物来源
  • K12 菌株

基因型

Δ(ara-leu) 7697 araD139  fhuA ΔlacX74 galK16 galE15 e14-  ϕ80dlacZΔM15  recA1 relA1 endA1 nupG  rpsL (StrR) rph spoT1 Δ(mrr-hsdRMS-mcrBC)

产品类别:
Cloning Competent Cell Strains Products

应用:
USER® Cloning,
Applications of USER® and Thermolabile USER II Enzymes,
Site Directed Mutagenesis,

Site Directed Mutagenesis,
High-throughput cloning and automation solutions,

Transformation

  • 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
    • C3019H     -80    
        NEB® 10-beta/Stable Outgrowth Medium B9035SVIAL 4 1 x 25 ml Not Applicable
        pUC19 Vector N3041AVIAL -20 1 x 0.025 ml 50 pg/µl
        NEB® 10-beta Competent E. coli (High Efficiency) C3019HVIAL -80 20 x 0.05 ml Not Applicable
    • C3019I     -80    
        NEB® 10-beta/Stable Outgrowth Medium B9035SVIAL 4 1 x 25 ml Not Applicable
        pUC19 Vector N3041AVIAL -20 1 x 0.025 ml 50 pg/µl
        NEB® 10-beta Competent E. coli (High Efficiency) C3019IVIAL -80 6 x 0.2 ml Not Applicable
    • C3019P     -80    
        pUC19 Vector N3041AVIAL -20 1 x 0.025 ml 50 pg/µl
        NEB® 10-beta/Stable Outgrowth Medium B9035SVIAL 4 1 x 25 ml Not Applicable
        NEB® 10-beta Competent E. coli (High Efficiency) C3019PVIAL -80 1 x 96-well plate
      (20 μl/well)
      Not Applicable

  • 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
    Tetracycline 15 µg/ml

    货运单

    • Ships on dry ice

    抗生素耐药性

    • str

  • 优势和特性

    Features

    • 克隆大质粒和 BAC
    • DH10B™ 衍生菌株

    应用特性

    质粒大小对转化效率的影响:10-beta E. coli 感受态细胞(高效级) |
    化学法制备的 NEB 10-beta 感受态细胞(C3019H)比 NEB 5-alpha 细胞(C2987H)更适合于大质粒的转化。质粒越大,两种感受态细胞的转化效率的差值就越大。
    热激时间对 NEB 10-beta E. coli 感受态细胞转化效率的影响:10-beta E. coli 感受态细胞(高效级) |  10-beta E. coli 感受态细胞(高效级) |按照所提供的高效级转化操作流程,用 100 pg pUC19 对照 DNA 转化 50 μl 感受态细胞(C3019H),仅热激时间从 0 至 80 秒不等。
    DNA 温育时间对 NEB 10-beta E. coli 感受态细胞转化效率的影响:10-beta E. coli 感受态细胞(高效级) |
    按照所提供的高效级转化操作流程,用 100 pg pUC19 对照 DNA 转化 50 μl 感受态细胞(C3019H),仅 DNA 温育时间为 0 至 40 分钟不等。
    细胞复苏培养基对转化效率的影响:10-beta E. coli 感受态细胞(高效级) |
    按照所提供的高效级转化操作流程,用 100 pg pUC19 对照 DNA 转化 50 μl 感受态细胞(C3019H),仅使用的复苏培养基不同。NEB 10-beta/Stable Outgrowth 培养基的转化效率最高。
    NEB 10-beta(C3019H/I)高转化效率带来的巨大优势:10-beta E. coli 感受态细胞(高效级) |
    使用不同厂家的产品,按照所提供的产品说明书操作,比较转化效率。图中数值取自三次平行实验的平均值。

     

  • 相关产品

    单独销售的组分

    • NEB® 10-beta/Stable Outgrowth 培养基

  • 注意事项

    1. 贮存和处理:感受态细胞应在 -80℃ 条件下贮存。-20℃ 条件下贮存会显著降低转化效率。当细胞温度高于 -80℃ 时,即使不融化,细胞也会失去效率。
    2. 注意:本产品含危险物质 DMSO。处理前请先查看 MSDS。

操作说明、说明书 & 用法

  • 操作说明

    1. High Efficiency Transformation Protocol using NEB 10-beta Competent E. coli (High Efficiency) (C3019H/C3019I)
    2. 5 Minute Transformation Protocol using NEB10-beta Competent E. coli  (C3019H/C3019I)
    3. High Efficiency Transformation Protocol with NEB 10-beta in 96-well Plate Format (NEB# C3019P)

  • 使用指南

    • Additional E. coli Strain Genotypes
    • Chemical Transformation Tips
    • Genetic Markers
    • McrA, McrBC and EcoKI Strain Phenotypes
    • Restriction of Foreign DNA by E. coli K-12

  • 应用实例

    • Enhancing Transformation Efficiency

工具 & 资源

  • 选择指南

    • Characteristics of Select E.coli Strains
    • Competent Cell Product Comparison
    • Competent Cell Selection Guide

  • Web 工具

    • Competitor Cross-Reference Tool
    • NEBcloner®

FAQs & 问题解决指南

  • FAQs

    1. Which competent cell strains are compatible with Gateway® Cloning?
    2. What are the strain properties (C3019)?
    3. What is the difference between NEB #C3019H and NEB #C3019I?
    4. What is the shelf life for this strain (NEB #C3019H and NEB #C3019I)?
    5. Which strain of Competent E.coli should I use for general cloning?
    6. Does plasmid size affect transformation efficiency (C3019)?
    7. How should I calculate the transformation efficiency (C3019)?
    8. Can I store competent cells at -20°C instead of -80°C?
    9. What is the optimal heat shock time for this strain (NEB #C3019H and NEB #C3019I)?
    10. How long should I incubate cells on ice after DNA has been added (NEB #C3019H and NEB #C3019I)?
    11. Which kind of transformation tubes should be used?
    12. What volume of DNA can be added into competent cells?
    13. Are NEB’s competent cells compatible with the “Mix & Go” protocol?
    14. What type of competent cells are suitable for transformation of DNA constructs created using NEBuilder HiFi DNA Assembly Master Mix?
    15. Why did Synthetic Biologist Chris Voigt of MIT choose NEB 10-beta for DNA assembly and cloning?
    16. How should I store the NEB 10-beta/Stable Outgrowth Medium?
    17. How should fragments be prepared for assembly using NEBuilder HiFi?
    18. Can the 96-well plate format of NEB 10-beta Competent E.coli, NEB #C3019P, be separated into smaller sections?
    19. How does the transformation efficiency of the 96-well plate format (NEB #C3019P) compare to the other formats?