NEB^® 10-beta Electrocompetent E. coli

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

Catalog #	Concentration	Size	List Price	Your Price
C3020K	Not Applicable	6 x 0.1 ml	$221.00	Sign In Sign In

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Product Information

NEB 10-beta 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

DH10B™ derivative
Transformation efficiency: > 2 x 10¹⁰ cfu/μg pUC19
Accomodation of large plasmids including BAC and cosmid constructs
Efficient transformation of methylated DNA derived from eukaryotic sources and unmethylated DNA derived from PCR, cDNA and other sources
Activity of nonspecific endonuclease I eliminated for highest quality plasmid preparations (endA1)
Suitable for blue/white screening by α-complementation of the β-galactosidase gene (Φ80ΔlacZM15)
Reduced recombination of cloned DNA (recA)
Resistance to phage T1 (fhuA)
K-12 Strain
Free of animal products

Genotype

Δ(ara-leu) 7697 araD139 fhuA ΔlacX74 galK16 galE15 e14- Φ80dlacZΔM15 recA1 relA1 endA1 nupG rpsL (Str^R) rph spoT1 Δ(mrr-hsdRMS-mcrBC)

Reagents Supplied

The following reagents are supplied with this product:

Show all

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

C3020K

-80

NEB^® 10-beta Electrocompetent E. coli	C3020KVIAL	-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

Product Categories:: Cloning Competent Cell Strains Products

Applications:: Transformation

Advantages and Features
Features
Electroporation Tips:
1. Electroporation cuvettes and microcentrifuge tubes should be pre-chilled on ice.
2. Electrocompetent cells should be thawed on ice and suspended well by carefully flicking the tubes.
3. 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.
4. 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 than10-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 (NEB #T1030) are recommended for cleanup of ligation reactions.
5. 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 1mm gap are recommended (e.g. BTX Model 610/613 and Bio-Rad #165-2089). Higher voltage is required for cuvettes with 2 mm gap.
6. Arcing may occur due to high concentration of salts or air bubbles.
7. It is essential to add recovery medium to the cells immediately after electroporation. One minute delay can cause a 3-fold reduction in efficiency.
8. Cold and dry selection plates lead to lower transformation efficiency. Pre-warm plates at 37°C for 1 hour. Using 37°C pre-warmed recovery medium increases the efficiency by about 20%.
9. 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.
Application Features

DNA Effects on Transformation Efficiency and Colony Output: Electroporation efficiency remains extremely high up to about 10 ng of input DNA, then decreases at higher DNA concentrations. Total colony output continues to increase with increasing DNA input up to at least 1 μg of puc19.
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
Companion Products
Materials Sold Separately
- NEB® 10-beta/Stable Outgrowth Medium
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.

Protocols, Manuals & Usage
- Protocols
  Electroporation Protocol (C3020)
- Usage Guidelines
  Additional E. coli Strain Genotypes
  
  Electroporation Tips
  
  Genetic Markers
  
  McrA, McrBC and EcoKI Strain Phenotypes
  
  Traditional Cloning Quick Guide
- 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.)
  
  C3020Datasheet-Lot40
  
  C3020Datasheet-Lot41
  
  C3020Datasheet-Lot44
  
  C3020Datasheet-Lot46
  
  C3020Datasheet-Lot50
  
  C3020Datasheet-Lot51
  
  C3020Datasheet-Lot52
  
  C3020Datasheet-Lot54
  
  C3020Datasheet-Lot0561405
  
  C3020Datasheet-Lot0551405
  
  C3020Datasheet-Lot0581406
  
  C3020Datasheet-Lot0581408
  
  C3020Datasheet-Lot0591408
  
  C3020Datasheet-Lot0601411
  
  C3020KDatasheet-Lot0601412
  
  C3020Datasheet-Lot0601501
  
  C3020KDatasheet-Lot0631504
Tools & Resources
- Selection Charts
  Characteristics of Select E.coli Strains
  
  Competent Cell Product Comparison
- Web Tools
  Competitor Cross-Reference Tool
  
  NEBcloner®
FAQs & Troubleshooting
- FAQs
  How should I calculate the Electrotransformation efficiency (C3020)?
  
  What are the strain properties (C3020)?
  
  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 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?
  
  How should I store the NEB 10-beta/Stable Outgrowth Medium?
  
  How should fragments be prepared for assembly using NEBuilder HiFi?
Citations & Technical Literature
- Citations
  
  Product Citation Tool
  
  Powered by Bioz See more details on Bioz
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
  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]
  
  C3020K_v1
  
  C3020S_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: [Product Number]_[Size]_[Version]_[Lot Number]
  
  C3020K_v1_0831701
  
  C3020K_v1_0851704
  
  C3020K_v1_0861704
  
  C3020K_v1_0861705
  
  C3020K_v1_0871706
  
  C3020K_v1_0871707
  
  C3020K_v1_0881707
  
  C3020K_v1_0891708
  
  C3020K_v1_0901710
  
  C3020K_v1_0921711
  
  C3020K_v1_0931712
  
  C3020K_v1_0931801
  
  C3020K_v1_0941802
  
  C3020K_v1_0951712
  
  C3020K_v1_0961803
  
  C3020K_v1_0971804
  
  C3020K_v1_10009785
  
  C3020K_v1_10012780
  
  C3020K_v1_10014832
  
  C3020K_v1_10016130
  
  C3020K_v1_10017214
  
  C3020K_v1_10019858
  
  C3020K_v1_10022925
  
  C3020K_v1_10024577
  
  C3020K_v1_10025364
  
  C3020K_v1_10026184
  
  C3020K_v1_10028860
  
  C3020K_v1_10029523
  
  C3020K_v1_10031152
  
  C3020K_v1_10032870
  
  C3020K_v1_10035090
  
  C3020K_v1_10035473
  
  C3020K_v1_10037874
  
  C3020K_v1_10041918
  
  C3020K_v1_10042809
  
  C3020K_v1_10044558
  
  C3020K_v1_10046069
  
  C3020K_v1_10047354
  
  C3020K_v1_10049960
  
  C3020K_v1_10051584
  
  C3020K_v1_10053711
  
  C3020K_v1_10055943
  
  C3020K_v1_10056800
  
  C3020K_v1_10057763
  
  C3020K_v1_10058619
  
  C3020K_v1_10060041
  
  C3020K_v1_10062923
  
  C3020K_v1_10063342
  
  C3020K_v1_10066623
  
  C3020K_v1_10068472
  
  C3020K_v1_10070090
  
  C3020K_v1_10070565
  
  C3020K_v1_10076055
  
  C3020K_v1_10077117
  
  C3020K_v1_10078740
  
  C3020K_v1_10080869
  
  C3020K_v1_10083573
  
  C3020K_v1_10085401
  
  C3020K_v1_10089108
  
  C3020K_v1_10089245
  
  C3020K_v1_10092564
  
  C3020S_v1_10096654
  
  C3020S_v1_10097074
  
  C3020K_v1_10102863
  
  C3020S_v1_10104339
  
  C3020K_v1_10105828
  
  C3020K_v1_10108886
  
  C3020K_v1_10109775
  
  C3020K_v1_10115183
  
  C3020K_v1_10118468
  
  C3020S_v1_10118882
  
  C3020K_v1_10123919
  
  C3020S_v1_10125368
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  NEB^® 10-beta Electrocompetent E. coli
  
  NEB^® 10-beta/Stable Outgrowth Medium
  
  pUC19 Vector
- 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.)
  
  C3020Datasheet-Lot40
  
  C3020Datasheet-Lot41
  
  C3020Datasheet-Lot44
  
  C3020Datasheet-Lot46
  
  C3020Datasheet-Lot50
  
  C3020Datasheet-Lot51
  
  C3020Datasheet-Lot52
  
  C3020Datasheet-Lot54
  
  C3020Datasheet-Lot0561405
  
  C3020Datasheet-Lot0551405
  
  C3020Datasheet-Lot0581406
  
  C3020Datasheet-Lot0581408
  
  C3020Datasheet-Lot0591408
  
  C3020Datasheet-Lot0601411
  
  C3020KDatasheet-Lot0601412
  
  C3020Datasheet-Lot0601501
  
  C3020KDatasheet-Lot0631504
- 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 contact NEB's Global Business Development team at gbd@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.