NEB Turbo Electrocompetent E. coli

• 产品编号：C2986K      品牌：New England Biolabs       原厂货号：C2986K
• 产品分类：克隆 > 克隆载体和克隆试剂盒 > 细菌菌株和感受态细胞 > 感受态细胞
• 应用分类：

描述：

• Transformation efficiency 1 x 10¹⁰cfu/µg pUC19 DNA
• Tight control of expression by lacI^q allows potentially toxic genes to be cloned
• Highest growth rate on agar plates - visible colonies 6.5 hours after transformation
• Activity of nonspecific endonuclease I (endA) eliminated for highest quality plasmid preparations
• Resistance to phage T1 (fhuA2)
• Suitable for blue/white screening by α-complementation of the β-galactosidase gene
• Isolate DNA after 4 hours of growth from a single overnight colony
• EcoKr^-m^-, McrBC^-
• K12 Strain

Description:
NEB Turbo Electrocompetent E. coli cells are suitable for high efficiency electrotransformation and rapid colony growth. These cells are ideal for DNA library constructions and all cloning purposes.

Reagents Supplied:
C2986K
6 x 0.1 ml/tube of NEB Turbo Electrocompetent E.coli cells (Store at -80°C)
25 ml of SOC Outgrowth Medium (Store at room temperature)
0.025 ml of 50 pg/µl pUC19 Control DNA (Store at -20°C)

Genotype: F' proA⁺B⁺ lacI^q ∆lacZM15/ fhuA2 ∆(lac-proAB) glnV gal R(zgb-210::Tn10)Tet^S endA1 thi-1 ∆(hsdS-mcrB)5

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 electroporation 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 are recommended for clean up 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 and/or increased resistance 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 of 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.
 

原厂资料：

注意事项：

General notes:

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.