NEB Stable Electrocompetent E. coli

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
C3041K     -80       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

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. 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.

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

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. Electroporation Protocol for NEB® Stable Electrocompetent E. coli (NEB #C3041K)

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

• Competitor Cross-Reference Tool

• NEBcloner^®

1. How should I calculate the electrotransformation efficiency (NEB #C3020/#C3041)?
2. What are the strain properties for NEB Stable (NEB #C3040/#C3041)
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 the NEB 10-beta/Stable Outgrowth Medium?

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 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]

• C3041K_v1_10346359

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• NEB Stable Electrocompetent E. coli
• NEB^® 10-beta/Stable Outgrowth Medium
• pUC19 Vector

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.

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