NEB Stable Electrocompetent E. coli
NEB Stable Electrocompetent E. coli
Electrocompetent E. coli cells suitable for high efficiency transformation.
- Ideal for isolation of plasmid clones containing repeat elements and unstable inserts
- Available in chemically competent format (single-use vials, 200 μl vials and 96-well plate)
- Useful for isolating and propagating retroviral/lentiviral clones
- Carries the endA mutation (in contrast to Stbl3) so that isolated plasmids are free of Endonuclease I
- No dry ice surcharge on competent cell shipments
- Outgrowth medium included
- Free of animal products
- Refer to our Electroporation Tips
Product Information
NEB Stable Electrocompetent E. coli cells are optimized for high efficiency transformation by electroporation. These cells are ideal for DNA library constructions and all cloning purposes.
Highlights
- Recommended host strain for cloning genes into retroviral/lentiviral vectors
- Recommended for cloning of direct repeats and inverted repeat sequences
- Reduced recombination of cloned DNA (recA1)
- 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 with vectors capable of alpha-complementation
Transformation Efficiency
Genotype
F' proA+B+ lacIq Δ(lacZ)M15 zzf::Tn10 (TetR) / Δ(ara-leu) 7697 araD139 fhuA ΔlacX74 galK16 galE15 e14- Φ80dlacZΔM15 recA1 relA1 endA1 nupG rpsL (StrR) rph spoT1 Δ(mrr-hsdRMS-mcrBC)
Product Categories
- Cloning Competent Cell Strains
Applications
- Transformation
Reagents Supplied
The following reagents are supplied with this product:
| Component Name | Component # | Stored at (°C) | Amount | Concentration |
|---|---|---|---|---|
| NEB Stable Electrocompetent E. coli | C3041KVIAL | -80 | 6 x 0.1 ml | Not Applicable |
| 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 |
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 |
Shipping Notes
- Ships on dry ice
Antibiotic Resistance
- tetracycline
- streptomycin
Advantages and Features
Features
Electroporation Tips
- Electroporation cuvettes and microcentrifuge tubes should be pre-chilled on ice.
- Electrocompetent cells should be thawed on ice and suspended well by carefully flicking the tubes.
- Once DNA is added to the cells, electroporation can be carried out immediately. It is not necessary to incubate DNA with cells. The maximum recommended volume of a DNA solution to be added is 2.5 µl. Addition of a large volume of DNA decreases transformation efficiency.
- Contaminants such as salts and proteins can lower electroporation efficiency. Ideally, DNA for transformation should be purified and suspended in water or TE. Transformation efficiency is more than 10-fold lower for ligation mixtures than the control pUC19 plasmid due to the presence of ligase and salts. If used directly, ligation reactions should be heat-inactivated at 65°C for 20 min and then diluted 10-fold. For optimal results, spin columns are recommended for cleanup of ligation reactions.
- Electroporation conditions vary with different cuvettes and electroporators. If you are using electroporators or cuvettes not specified in the protocol, you may need to optimize the electroporation conditions. Cuvettes with 1 mm gap are recommended (e.g. BTX Model 610/613 and Bio-Rad #165-2089). Higher voltage is required for cuvettes with 2 mm gap.
- Arcing may occur due to high concentration of salts or air bubbles.
- It is essential to add recovery medium to the cells immediately after electroporation. One minute delay can cause a 3-fold reduction in efficiency.
- Refreeze unused cells in a dry ice/ethanol bath for 5 min and then store at -80°C. Do not use liquid nitrogen. Additional freeze-thaw cycles result in lower transformation efficiency.
Related Products
Companion Products
- ElectroLigase®
- NEB® Stable Competent E. coli (High Efficiency)
- NEB® 10-beta Electrocompetent E. coli
- NEB Tube Opener
Materials Sold Separately
- NEB® 10-beta/Stable Outgrowth Medium
Product Notes
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 for this Product
- Electroporation Protocol for NEB® Stable Electrocompetent E. coli (NEB #C3041K)
Usage Guidelines
- Additional E. coli Strain Genotypes
- Electroporation Tips
- Genetic Markers
- McrA, McrBC and EcoKI Strain Phenotypes
- Traditional Cloning Quick Guide
Tools & Resources
Web Tools
- Competitor Cross-Reference Tool
- NEBcloner®
FAQs & Troubleshooting
FAQs
- How should I calculate the electrotransformation efficiency (NEB #C3020/#C3041)?
- What are the strain properties for NEB Stable (NEB #C3040/#C3041)?
- What type of competent cells are suitable for transformation of DNA constructs created using Gibson Assembly?
- What type of competent cells are suitable for transformation of DNA constructs created using NEBuilder HiFi DNA Assembly Master Mix?
- How should I store the NEB 10-beta/Stable Outgrowth Medium?
Quality, Safety & Legal
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.
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:
- C3041K_v1
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:
- C3041K_v1_10346359
- C3041K_v1_10353328
- C3041K_v1_10357167
Safety Data Sheets
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use the product safely.
- NEB Stable Electrocompetent E. coli
- NEB® 10-beta/Stable Outgrowth Medium
- pUC19 Vector
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.
