NEB® 10-beta Electrocompetent E. coli

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 electroporator. If you are using electroporators 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

Companion Products

• NEB® Turbo Electrocompetent E. coli
• NEB® 5-alpha Electrocompetent E. coli
• NEB® 10-beta Competent E. coli (High Efficiency)
• phi29 DNA Polymerase
• ElectroLigase®
• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)

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.
   

1. How should I calculate the Electrotransformation efficiency (C3020)?
2. What are the strain properties (C3020)?
3. What type of competent cells are suitable for transformation of DNA constructs created using Gibson Assembly?
4. What type of competent cells are suitable for transformation of DNA constructs created using NEBuilder HiFi DNA Assembly Master Mix?
5. 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?
6. How should I store the NEB 10-beta/Stable Outgrowth Medium?

1. Electroporation Protocol (C3020)

• Characteristics of Select E.coli Strains
• Competent Cell Product Comparison

• Additional E. coli Strain Genotypes
• Electroporation Tips
• Genetic Markers
• McrA, McrBC and EcoKI Strain Phenotypes
• Traditional Cloning Quick Guide

• Competitor Cross-Reference Tool

• NEBcloner

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.

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

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

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

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 [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.