Product Class: Other

NEB® 5-alpha Competent E. coli (High Efficiency)

Also available in Electrocompetent format.

Product Introduction

NEB 5-alpha Competent E. coli is a derivative of the popular DH5α. It is T1 phage resistant and endA deficient for high-quality plasmid preparations. 

  • High efficiency strain ideal for a wide variety of applications
  • Available in a wide variety of sizes, including single-use vials, 200 μl vials, 96- and 384- well plate and 12 x 8 tube strips
  • No dry ice surcharge on competent cell shipments
  • Outgrowth medium included
  • Free of animal products
Catalog # Size Concentration
C2987H 20 x 0.05 ml/tube 0 mg/ml
C2987I 6 x 0.2 ml/tube 0 mg/ml
C2987P 1 x 96 well plate (20 µl/well) 0 %
C2987R 1 x 384 well plate (10 µl/well) 0 %
C2987U 96 x 50 μl/tube (12 x 8-tube strips) 0 %

Product Information

Description

Highlights

  • DH5α™ derivative
  • Free of animal products
  • Transformation efficiency:
    For C2987H, C2987I, C2987P and C2987U: 1 - 3 x 109 cfu/μg pUC19 DNA
    For C2987R: 1 - 5 x 108 cfu/μg pUC19 DNA
  • Efficient transformation of unmethylated DNA derived from PCR, cDNA and many other sources (hsdR)
  • Activity of nonspecific endonuclease I (endA1) eliminated for highest quality plasmid preparations
  • Resistance to phage T1 (fhuA2)
  • Suitable for blue/white screening by α-complementation of the β-galactosidase gene
  • Reduced recombination of cloned DNA (recA1)
  • K12 Strain

Chemically competent E. coli cells suitable for high efficiency transformation in a wide variety of applications. Available in 5 convenient formats:

 

For C2987H, C2987I, C2987P and C2987U, SOC Outgrowth Medium is provided.
For C2987R, Outgrowth Medium 1.5 is provided.

Genotype

fhuA2 Δ(argF-lacZ)U169 phoA glnV44 Φ80 Δ(lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
SOC Outgrowth Medium 4 1 X
pUC19 Vector -20 0.05 ng/μl
Product Categories:
Cloning Strains
Applications:
USER® Cloning,
Transformation,
Applications of USER® and Thermolabile USER II Enzymes

Advantages and Features

Features

  • Versatile cloning strain
  • DH5α™ derivative

Application Features


DNA Effects on Transformation Efficiency and Colony Output: DNA Effects on Transformation Efficiency and Colony Output:
The optimal amount of DNA to use in a transformation reaction is lower than commonly recognized. Using clean, supercoiled pUC19, the efficiency of transformation is highest in the 100 pg-1 ng range. However, the total colonies which can be obtained from a single transformation reaction increase up to about 100 ng.
Effect of heat shock time on NEB 5-alpha competent E.coli transformation efficiency: Effect of heat shock time on NEB 5-alpha 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 5-alpha competent E.coli transformation efficiency: Effect of DNA incubation time on NEB 5-alpha 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: Effect of outgrowth medium on transformation efficiency:
50 μl of NEB 5-alpha 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.
Benefit from the high transformation efficiencies of NEB 5-alpha: Benefit from the high transformation efficiencies of NEB 5-alpha:
The transformation efficiencies of NEB 5-alpha and DH5α were compared using each manufacturers' recommended protocols. Values shown are the average of triplicate experiments.

Properties and Usage

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
Streptomycin 25 μg/ml

Storage Temperature

-80°C

Shipping Notes

  • Ships on dry ice

Antibiotic Resistance

  • none

Notes

  1. For C2987H, C2987I, C2987P and C2987U, SOC Outgrowth Medium is provided.
    For C2987R, Outgrowth Medium 1.5 is provided.

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

  3. 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.

    SOC medium can be stored at either 4°C or Room Temperature depending on how fast it will be used. Storing at Room Temperature is convenient and adequate for short term usage (weeks to a couple of months). For long term storage, we recommend storing at 4°C. Please note that Outgrowth Medium 1.5 supplied with C2987R (1 x 384 well plate format) must only be stored at Room Temperature or crystals will form.

Faqs & Tech Tips

FAQs

  1. Which competent cell strains are compatible with Gateway® Cloning?  Which strains are not compatible?
  2. Can I store competent cells at -20°C instead of -80°C?
  3. Do you carry competent cells which are compatible with Gateway® Cloning?
  4. How can I increase transformation efficiency?
  5. How long should I incubate cells on ice after DNA has been added (NEB #C2987H and NEB #C2987I)?
  6. How should I calculate the transformation efficiency (C2987)?
  7. What are the solutions/recipes (C2987)?
  8. What are the strain properties (C2987)?
  9. What is the difference between NEB #C2987H and NEB #C2987I?
  10. What is the difference between NEB #C2988J and NEB #C2987H?
  11. What is the optimal heat shock time for this strain (NEB #C2987H and NEB #C2987I)?
  12. What type of competent cells are suitable for transformation of DNA constructs created using Gibson Assembly?
  13. Which strain of Competent E. coli should I use for general cloning?
  14. Which kind of transformation tubes should be used?
  15. What volume of DNA can be added into competent cells?
  16. Are NEB's competent cells compatible with the “Mix & Go" protocol?
  17. What is the shelf life for this strain (NEB #C2987H and NEB #C2987I)?
  18. Can the 96-well plate format of NEB 5-alpha Competent E.coli, NEB #C2987P, be separated into smaller sections?
  19. How does the transformation efficiency of the 96-well plate format (NEB #C2987P) compare to the other formats?
  20. What is the optimal heat shock time for the 96-well plate format NEB 5-alpha Competent E.coli (NEB #C2987P)?
  21. I am competing in the iGEM competition. Do you have any products that I should consider from NEB?
  22. What is the optimal heat shock time for the 384-well plate format NEB 5-alpha Competent E. coli (NEB #C2987R)?
  23. What is the optimal heat shock time for the 96-tube format NEB 5-alpha Competent E. coli (NEB #C2987U)?
  24. What type of competent cells are suitable for transformation of DNA constructs created using NEBuilder HiFi DNA Assembly Master Mix?
  25. 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?

Protocols & Manuals

Protocols

  1. 5 Minute Transformation Protocol (C2987H/C2987I)
  2. High Efficiency Transformation Protocol (C2987H/C2987I)
  3. High Efficiency Transformation Protocol for 96-well format (C2987P)
  4. 14 Minute Transformation Protocol (C2987H/C2987I)
  5. High Efficiency Transformation Protocol for 96-tube format (C2987U)
  6. High Efficiency Transformation Protocol for 384-well format (C2987R)

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.)

Other Tools & Resources

Selection Charts

Usage Guildelines & Tips

Interactive Tools

Application Notes

Citations

  • Lee S.B., Park S.K., Kim Y.S. (2015) Production of bioactive chicken (Gallus gallus) follistatin-type proteins in E. coli Sci Rep; 5, 11603. PubMedID: 26302688 , DOI: 10.1038/srep11603
  • Kostylev M., Otwell A.E., Richardson R., Suzuki Y. (2015) Cloning Should Be Simple: Escherichia coli DH5α-Mediated Assembly of Multiple DNA Fragments with Short End Homologies PLoS One; 10, e0137466. PubMedID: PMC4562628 , DOI: 10.1371/journal.pone.0137466
  • Mascher G, Derman Y, Kirk DG, Palonen E, Lindström M, Korkeala H (2014) The CLO3403/CLO3404 Two-Component System of Clostridium botulinum E1 Beluga Is Important for Cold Shock Response and Growth at Low Temperatures Appl Environ Microbiol; 80(1), 399-407. PubMedID: 24185852 , DOI: 10.1128/AEM.03204-13

Quality & Safety

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 control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]