NEB 5-alpha Competent E. coli (Subcloning Efficiency)

• 产品编号：C2988J      品牌：New England Biolabs       原厂货号：C2988J
• 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：

Description:
Chemically competent E. coli cells suitable for subcloning efficiency transformation in a wide variety of applications.

Genotype: fhuA2 Δ(argF-lacZ)U169 phoA glnV44 Φ80Δ (lacZ)M15 gyrA96 recA1 relA1 endA1 thi-1 hsdR17

Reagents Supplied:
6 x 0.4 ml/tube of chemically competent NEB 5-alpha Competent E. coli cells (Store at -80°C)

Transformation Protocol Variables:
Thawing: Cells are best thawed on ice and DNA added as soon as the last bit of ice in the tube disappears. Cells can also be thawed by hand, but warming above 0°C will decrease the transformation efficiency.

Incubation of DNA with Cells on Ice: For maximum transformation efficiency, cells and DNA should be incubated together on ice for 30 minutes. Expect a 2-fold loss in transformation efficiency for every 10 minutes this step is shortened.

Heat Shock: Both the temperature and the timing of the heat shock step are important and specific to the transformation volume and vessel. Using the transformation tube provided, 30 seconds at 42°C is optimal.

Outgrowth: Outgrowth at 37°C for 1 hour is best for cell recovery and for expression of antibiotic resistance. Expect a 2-fold loss in transformation efficiency for every 15 minutes this step is shortened. SOC gives 2-fold higher transformation efficiency than LB medium; and incubation with shaking or rotating the tube gives 2-fold higher transformation efficiency than incubation without shaking.

Plating: Selection plates can be used warm or cold, wet or dry without significantly affecting the transformation efficiency. However, warm, dry plates are easier to spread and allow for the most rapid colony formation.
 

原厂资料：

注意事项：

CAUTION: This product contains DMSO, a hazardous material. Review the MSDS before handling.

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.