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  • NEB 10-beta Competent E. coli (High Efficiency)

    Description

    Chemically competent E. coli cells suitable for high efficiency transformation in a wide variety of applications.

    Highlights

    • Transformation efficiency: 1–3 x 109 cfu/µg pUC19 DNA
    • Efficient transformation of methylated DNA derived from eukaryotic sources or unmethylated DNA derived from PCR, cDNA and many other sources [mcrAΔ(mrr-hsdRMS-mcrBC)]
    • Activity of nonspecific endonuclease I (endA1) eliminated for highest quality plasmid preparations
    • Resistance to phage T1 (fhuA2)
    • Suitable for blue/white screening without IPTG by a-complementation of the b-galactosidase gene
    • Reduced recombination of cloned DNA (recA1)
    • Clone large plasmids and BACs
    • DH10B™ derivative
    • Free of animal products
    • K12 Strain

    Genotype

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

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    pUC19 Transformation Control Plasmid-200.05 ng/μl
    SOC Outgrowth Medium41X

    Advantages and Features

    Features

    • Large plasmid and BAC cloning
    • DH10B™ derivative

    Applications


    Effect of Plasmid Size on Transformation Efficiency: NEB 10-beta chemically competent cells are more efficiently transformed with large plasmids than NEB 5-alpha cells. The difference in TE between the two cell lines increases with the size of the plasmid being transformed.
    Effect of heat shock time on NEB 10-beta competent E. coli transformation efficiency: 50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except heat shock time varied from 0 to 80 seconds.
    Effect of DNA incubation time on NEB 10-beta competent E. coli transformation efficiency: 50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except DNA incubation time varied from 0 to 40 minutes.
    Effect of outgrowth medium on transformation efficiency: 50 μl of NEB 10-beta competent E. coli was transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol with the exception of varying the outgrowth medium. NEB SOC outgrowth medium delivers the highest transformation efficiency.
    Take advantage of the low cost per transformation with NEB 10-beta. Calculations were based on list price and recommended transformation volumes.

    Properties and Usage

    Antibiotics for Plasmid SelectionWorking Concentration
    Ampicillin100 μg/ml
    Carbenicillin100 μg/ml
    Chloramphenicol33 μg/ml
    Kanamycin30 μg/ml
    Tetracycline15 μg/ml

    Storage Temperature

    -80°C

    Shipping Notes

    • Ships on dry ice

    Antibiotic Resistance

    • str

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Transformation Efficiency:
      The competent cells are tested for transformation efficiency and pass minimum release criteria. Transformation efficiency is defined as the number of colony forming units (cfu) which would be produced by transforming 1 μg of plasmid into a given volume of competent cells.

    Notes

    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.
    2. CAUTION: This product contains DMSO, a hazardous material. Review the MSDS before handling.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    1. Do you carry competent cells which are compatible with Gateway® Cloning?
    2. What are the solutions/recipes (C3019)?
    3. What are the strain properties (C3019)?
    4. What is the difference between NEB #C3019H and NEB #C3019I?
    5. What is the shelf life for this strain (NEB #C3019H and NEB #C3019I)?
    6. Which strain of Competent E. coli should I use for general cloning?
    7. Does plasmid size affect transformation efficiency (C3019)?
    8. How should I calculate the transformation efficiency (C3019)?
    9. Can I store competent cells at -20°C instead of -80°C?
    10. What is the optimal heat shock time for this strain (NEB #C3019H and NEB #C3019I)?
    11. How long should I incubate cells on ice after DNA has been added (NEB #C3019H and NEB #C3019I)?
    12. Which kind of transformation tubes should be used?
    13. What volume of DNA can be added into competent cells?
    14. Are NEB's competent cells compatible with the "Plate and Go" protocol?
    1. High Efficiency Transformation Protocol using NEB 10-beta Competent E. coli (High Efficiency) (C3019)
    2. 5 Minute Transformation Protocol (C3019)

    Selection Tools

    Usage Guidelines & Tips

    Application Notes