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| | <noinclude>{{:Team:EPF-Lausanne/Template/Header}}</noinclude> | | <noinclude>{{:Team:EPF-Lausanne/Template/Header}}</noinclude> |
| - | {{:Team:EPF-Lausanne/Template/ProtocolHeader|E.coli Transformation|{{{1|}}}}} | + | |
| | + | <noinclude>{{:Team:EPF-Lausanne/Template/SetTitle|Transformation}}</noinclude> |
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| | + | {{:Team:EPF-Lausanne/Template/ProtocolHeader|E.Coli Transformation|{{{1|}}}}} |
| | <!-- /\ --> | | <!-- /\ --> |
| | <!-- || --> | | <!-- || --> |
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| | # Thaw the competent E.coli (DH5alpha) cells on ice (not in hands!) | | # Thaw the competent E.coli (DH5alpha) cells on ice (not in hands!) |
| - | # As soon as it is thawed, add 50ul of the cells to the DNA (~50-100ng of pure plasmid) | + | # As soon as it is thawed, add 50µl of the cells to the DNA (~50-100 ng of pure plasmid, or some 2 µl usually) |
| - | # Let it rest on ice for 20-30 min. Meanwhile, put agar plate (with correct antibiotic) | + | # Let it rest on ice for 20-30 min. Meanwhile, put agar plate (with correct antibiotic) at 37°C for prewarming. |
| - | at 37°C for prewarming. | + | # Put the tube with DNA+E.coli at 42°C for 45 sec - 1 min (heat shock) |
| - | # Put the tube with DNA+E.coli at 42°C for 45 sec-1 min (heat shock) | + | # Add 400 µl of LB broth and place at 37°C for 20-30 min (shaking) |
| - | # Add 400ul of LB broth and place at 37°C for 20-30min (shaking) | + | |
| | # Spread the cells on the prewarmed plate (and let it dry) | | # Spread the cells on the prewarmed plate (and let it dry) |
| - | # Incubate the plate upside-down at 37°C for ~14-15h (leaving it more than 16h decreases | + | # Incubate the plate upside-down at 37°C for ~14-15 hours (leaving it more than 16h decreases the plasmid quality) |
| - | the plasmid quality) | + | |
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| | {{:Team:EPF-Lausanne/Template/ProtocolFooter}} | | {{:Team:EPF-Lausanne/Template/ProtocolFooter}} |
| | <noinclude>{{:Team:EPF-Lausanne/Template/Footer}}</noinclude> | | <noinclude>{{:Team:EPF-Lausanne/Template/Footer}}</noinclude> |
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