K-12 MG1655 感受态细胞(化转)/ K-12 MG

K-12 MG1655 Chemically Competent Cell

Packing 10 x 0.1 ml/tube

Store at –80°C

 

Genotype
F- lambda- ilvG- rfb-50 rph-1(NC_000913)

Alternative name
K-12 MG1655 chemically Competent Cells,K-12 MG1655 Competent E.coli,K-12 MG1655 Competent Cells, K-12 MG1655 chemically E.coli Express Competent Cells 

Transformation Protocol

1.Please store K-12 MG1655 Chemically Competent Cell in -80°C for late use.

2. Thaw a tube of K-12 MG1655 Chemically Competent Cell锛?00μl 锛塷n ice.

3. Add 1µl plasmid DNA or 10µl ligation product to the cell mixture. Carefully flick the tube 4–5 times to mix cells and DNA. Do not vortex.

4.Place the mixture on ice for 30 minutes. Do not mix.

5. Heat shock at exactly 42°C for exactly 90 seconds and then immediately transfer the tube back into ice for 5min. Do not mix.

6. Place on ice for 5 minutes. Do not mix.

7. Add 800ul LB then put on the Constant temperature shaker (150 rpm) for 45min,Warm selection plates to 37°C.

8. Pellet the mixture by centrifugation at 4000rpm/min for 3min. Pour off about 200ul of cell supernatant, then resuspend the cell pellet in the remaining medium by gently vortexing the tube.

9.Spread the cell on the agar plates by antibiotic, then place it on 37°C for 1hour.

10. Incubate overnight at 37°C.

 

Caution:

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.

2. This product is FOR RESEARCH USE ONLY!