Protocol : Transformation of bacteria super competent cells

1. Thaw bacteria super competent cells at room temperature.

2. Control T-: In a 1.5 ml microcentrifuge tube (eppendorf) (0), add 100 μl solution of bacteria super competent cells.

Plasmid DNA Puc 18: In a 1.5 ml eppendorf (1), add 100 μl solution of bacteria super competent cells, 1 μl (concentration 0.1 ng/μl) d’AND Puc 18 (Resistant of Ampicillin).

3. Incubate solutions at 0 °C for 30 min.

4. Make a heat shock: incubate solutions at 42 °C for 1 min, then at 0 °C for 2 min.

5. Add 1 ml LB in two eppendorfs, incubate solutions with agitation at 37 °C for 1 h.

6. Dilution for the solution (1): In a 1.5 ml eppendorf (2), add 100 μl solution (1) and 900 μl LB. In a 1.5 ml eppendorf (3), add 100 μl solution (2) and 900 μl LB.

7. Take 100 μl of each solution, spread them out on four identical environment with ampicillin.