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Team:Paris Saclay/Protocols/Transformation
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In a 1.5 ml microcentrifuge tube (eppendorf) (0), add 100 μl solution of bacteria super competent cells. | In a 1.5 ml microcentrifuge tube (eppendorf) (0), add 100 μl solution of bacteria super competent cells. | ||
| - | Plasmid DNA | + | Plasmid DNA pUC18: |
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). | 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). | ||
Revision as of 14:40, 23 September 2013
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 pUC18: 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.
