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| - | ='''Protocol : Transformation of supercompetent ''E.coli'' cells'''=
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| - | # Thaw supercompetent cells at room temperature. (almost 100µL for control and 100 µL for one DNA transformation )
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| - | # Control T-:
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| - | In a 1.5 ml microcentrifuge tube (eppendorf), add 100 μl solution of supercompetent cells (0).
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| - | Plasmid DNA pUC18 (or DNA of your choice) :
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| - | In a 1.5 ml eppendorf , add 100 μl solution of supercompetent cells, 1 μl (concentration 0.1 ng/μl) of DNA pUC18 (Resistant of Ampicillin)(1).
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| - | # Incubate solutions on ice for 30 min.
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| - | 4. Make a heat shock: incubate bacteria at 42 °C for 1 min, then on ice for 2 min.
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| - | 5. Add 1 ml LB in two eppendorfs, incubate solutions with agitation at 37 °C for 1 h.
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| - | 6. Dilution for the solution (1): ( you are going to plate different dilutions of your pUC18 transformed bacteria)
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| - | * 10-1: In a 1.5 ml eppendorf (2), add 100 μl solution (1) and 900 μl LB.
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| - | * 10-2: In a 1.5 ml eppendorf (3), add 100 μl solution (2) and 900 μl LB.
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| - | 7. Take 100 μl of each solution : the control(0)and the pUC18 transformed bacteria (1),(2),(3) spread them out on four identical environment with ampicillin ( or appropriated antibiotic).
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| - | [[File:Pstransformation.jpg|center|200px]]
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Latest revision as of 10:01, 12 July 2014
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