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  2. Products  /  NEB CATALOGUE  /  Competent Cells (Cloning)  /  NEB<sup>®</sup> 10-beta Competent <em>E. coli</em> (High Efficiency)

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

#C3019

Catalog # Size Price (£)
C3019H 20 X 0.05 ML £198.00
C3019P 1 96-WELL PLATE £500.00
C3019I 6 X 0.2 ML £165.00

Need a custom/large volume order?  Contact Us

NEB 10-beta Competent E. coli is a derivative of the popular DH10B. It is T1 phage resistant and endonuclease I (endA1) deficient for high- quality plasmid preparations.

  • High efficiency strain ideal for cloning large plasmids and BACs

  • Available in single-use vials, 200 μl vials, 96-well plate and electrocompetent format

  • No dry ice surcharge on competent cell shipments

  • Outgrowth medium included

  • Free of animal products

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

Highlights

  • DH10B™ derivative

  • Transformation efficiency:
         For C3019I and C3019H:  1-3 x 109 cfu/µg pUC19 DNA
         For C3019P: 1-3 x 108 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 (fhuA)

  • 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

  • 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:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

  • 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

Advantages and Features

Features

  • Large plasmid and BAC cloning

  • DH10B™ derivative

Application Features


Effect of Plasmid Size on Transformation Efficiency: Effect of Plasmid Size on Transformation Efficiency: NEB 10-beta chemically competent cells (C3019H) are more efficiently transformed with large plasmids than NEB 5-alpha cells (C2987H). 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: Effect of heat shock time on NEB 10-beta competent E. coli transformation efficiency:

   50 μl of competent cells (C3019H) 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:  Effect of DNA incubation time on NEB 10-beta competent E. coli transformation efficiency: 50 μl of competent cells (C3019H) 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: Outgrowth 50 μl of NEB 10-beta competent E. coli (C3019H) 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 10-beta/Stable Outgrowth Medium delivers the highest transformation efficiency.

Benefit from the high transformation efficiencies of NEB 10-beta (C3019H/I): Efficiency Transformation efficiencies were compared using manufacturers' recommended protocols. Values shown are the average of triplicate experiments. 

 



Properties & Usage

Antibiotic for Plasmid Selection

Antibiotics for Plasmid Selection

Working Concentration

Ampicillin

100 µg/ml

Carbenicillin

100 µg/ml

Chloramphenicol

33 µg/ml

Kanamycin

30 µg/ml

Tetracycline

15 µg/ml

Shipping Notes

  • Ships on dry ice

Antibiotic Resistance

  • str

Related Products

Materials Sold Separately

Product 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.

Protocols

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

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.)

Quality Control Assays

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Certificate of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Specifications & Change Notifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

The following link is to a PDF Safety Data Sheet (SDS) that applies to this product to help you use it safely.



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.)

Legal and Disclaimers

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please email us at busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science - we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

Trademarks

ONESHOT® and MAX EFFICIENCY® are registered trademarks of Thermo Fisher Scientific.
DH10B™ is a trademarks of Thermo Fisher Scientific.



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