Product Class: Other

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

Supplied with outgrowth medium optimized for NEB 10-beta & NEB Stable Competent E.coli ; please do not use SOC outgrowth medium.

Now available in 96-well plate format.

Product Introduction

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
Catalog # Size Concentration
C3019H 20.0 x 0.05 ml
C3019I 6.0 x 0.2 ml

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Product Information

Description

Highlights

  • DH10B™ derivative
  • 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

Transformation Efficiency

For C3019I and C3019H:  1–3 x 109 cfu/µg pUC19 DNA
For C3019P: 1–3 x 108 cfu/µg pUC19 DNA 

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

Genotype

Δ(ara-leu) 7697 araD139  fhuA ΔlacX74 galK16 galE15 e14-  ϕ80dlacZΔM15  recA1 relA1 endA1 nupG  rpsL (StrR) rph spoT1 Δ(mrr-hsdRMS-mcrBC)
This product is related to the following categories:
Cloning Competent Cell Strains Products,
This product can be used in the following applications:
USER® Cloning,
Applications of USER® and Thermolabile USER II Enzymes,
Site Directed Mutagenesis,
Site Directed Mutagenesis,
Transformation,
High-throughput cloning and automation solutions

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

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

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.

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, Manuals & Usage

Protocols

  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)

Usage & Guidelines

Application Notes

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

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?

Quality, Safety & Legal

Quality Assurance Statement

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.

Specifications

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]

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]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email 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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