Product Class: Other

NEB® Turbo Competent E. coli (High Efficiency)

Product Introduction

E. coli cells featuring fast colony growth (6.5 hours)

  • Tight expression control (lacIq)
  • Isolate DNA after only 4 hours of growth
  • 5-minute transformation protocol with AmpR plasmids
  • No dry ice surcharge on competent cell shipments
  • Outgrowth medium included
  • Free of animal products
Catalog # Size Concentration
C2984H 20.0 x 0.05 ml
C2984I 6.0 x 0.2 ml

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Product Information

Description

Highlights

  • Tight control of expression by laclq allows potentially toxic genes to be cloned
  • Highest growth rate on agar plates - visible colonies 6.5 hours after transformation
  • Activity 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
  • Suitable for 5 minute transformation protocol with AmpR plasmids
  • EcoKr-m-, McrBC-
  • K12 Strain
  • Isolate DNA after 4 hours of growth from a single overnight colony
  • Free of animal products

Transformation Efficiency

1 - 3 x 109 cfu/μg pUC19 DNA Chemically competent E. coli cells suitable for high efficiency transformation and rapid colony growth.

Genotype

F' proA+B+ lacIq ∆lacZM15 / fhuA2  ∆(lac-proAB)  glnV galK16 galE15  R(zgb-210::Tn10)TetS  endA1 thi-1 ∆(hsdS-mcrB)5  

This product is related to the following categories:
Cloning Competent Cell Strains Products
This product can be used in the following applications:
High-throughput cloning and automation solutions,
Transformation,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB

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
Streptomycin 25 µg/ml
Tetracycline 15 µg/ml

Shipping Notes

  • Ships on dry ice

Antibiotic Resistance

  • nit

Features

  • Fastest growth – colonies visible after 6.5 hours
  • Plasmid preparation after 4 hours
  • Tight control/expression of toxic genes (F´lacIq)

Application Features


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.
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.

Growth curve of NEB Turbo vs DH5α at 37°C in shaking flask:Growth curve of NEB Turbo vs DH5α at 37°C in shaking flask:
NEB Turbo and DH5α frozen stock (same cell density) were inoculated 1:100 into 80 ml of liquid medium (10g tryptone, 5g yeast extract, 10g NaCl, 1g MgCl2 and 1g Dextrose per liter, pH7.2) in 1 liter flask, respectively. Cells were grown at 37°C with shaking at 250 rpm. O.D.600 was measured every 30 minutes for up to 16 hours.
Growth curve of NEB Turbo vs DH5α at 37°C in fermentor: Growth curve of NEB Turbo vs DH5α at 37°C in fermentor
NEB Turbo and DH5α cultures grown overnight were inoculated 1:50 into 1 liter of LB (10g tryptone, 5g yeast extract, 10g NaCl per liter, pH7.0) in 1 liter fermentor, respectively. Cells were grown at 37°C. O.D.600 was measured every 30 minutes for up to 9 hours.
NEB Turbo cell growth and DNA yield: NEB Turbo cell growth and DNA yield:
pUC19 was transformed into NEB Turbo competent E.coli. Six colonies grown overnight (colony size was about 2.4 mm in diameter) were inoculated into 30 ml of LB/Amp, mixed thoroughly and 5 ml was dispensed into each 25 ml test tube (equal to 1 colony in 5 ml of LB/Amp). The cells were grown at 37°C with shaking at 250 rpm. One test tube was placed on ice after 3, 4, 5, 6, 7 and 16 hours of growth, respectively and the O.D.600 was measured. Mini-prep was performed by spinning down 1.5 ml of culture 3 times (total 4.5 ml) and finally eluting the DNA into 50 μl of EB buffer.
Comparison of DNA yield after 4 hours of growth: Comparison of DNA yield after 4 hours of growth:
pUC19 was transformed into NEB Turbo competent E.coli, Invitrogen Mach1 and DH5α, respectively. One overnight grown colony (colony size for NEB Turbo was about 2.4 mm in diameter, 1.8 mm for Mach1 and 1.2 mm for DH5α) was inoculated into 5 ml of LB/Amp in a 25ml test tube and cells were grown at 37°C with shaking at 250 rpm. After 4 hours of growth, cells were measured O.D.600 and mini-prep was performed by spinning down 1.5 ml of culture 3 times (total 4.5 ml) and finally eluting the DNA into 50 μl of EB buffer. O.D.600 for NEB Turbo was 3.08, 2.36 for Mach1 and 0.75 for DH5α.

Effect of heat shock time on NEB Turbo competent E.coli transformation efficiency:Effect of heat shock time on NEB Turbo 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 Turbo competent E.coli transformation efficiency:Effect of DNA incubation time on NEB Turbo 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 Turbo 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.

Product Notes

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

Protocols, Manuals & Usage

Protocols

  1. High Efficiency Transformation Protocol (C2984)
  2. 5 Minute Transformation Protocol (C2984)

Usage & Guidelines

Application Notes

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Which competent cell strains are compatible with Gateway® Cloning?
  2. Does plasmid size affect colony size (C2984)?
  3. How long can I purify plasmid after the inoculation from a single colony using NEB Turbo competent E.coli (C2984)?
  4. How long should I incubate cells on ice after DNA has been added (NEB #C2984H and NEB #C2984I)?
  5. What are the strain properties (C2984)?
  6. What does the growth curve of NEB Turbo cells look like compared to DH5α?
  7. What is the difference between NEB #C2984H and NEB #C2984I?
  8. What is the optimal heat shock time for this strain (NEB #C2984H and NEB #C2984I)?
  9. Which strain of Competent E.coli should I use for general cloning?
  10. How should I calculate the transformation efficiency (C2984)?
  11. What are the solutions/recipes (C2984)?
  12. Can I store competent cells at -20°C instead of -80°C?
  13. Which kind of transformation tubes should be used?
  14. What volume of DNA can be added into competent cells?
  15. What is the shelf life for this strain (NEB #C2984H and NEB #C2984I)?
  16. Are NEB's competent cells compatible with the “Mix & Go" protocol?
  17. 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?
  18. How should fragments be prepared for assembly using NEBuilder HiFi?
  19. Are NEB Turbo Competent E. coli (High Efficiency) or NEB Turbo Electrocompetent E. coli recommended for cloning repetitive or unstable DNA?

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 [email protected].

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.

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