Transformation
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<b>Procedure</b> | <b>Procedure</b> | ||
| - | Preparation: competent cells | + | Preparation: [[competent cells]] |
For each plasmid: | For each plasmid: | ||
* take competent cells from -80C freezer | * take competent cells from -80C freezer | ||
| - | * | + | * thaw on ice for 10min |
* add 2uL DNA to competent cell (label) | * add 2uL DNA to competent cell (label) | ||
* mix with pipette | * mix with pipette | ||
| - | * sit for 30min | + | * sit on ice for 30min |
* get a beaker of exactly 42C water from hot water tap | * get a beaker of exactly 42C water from hot water tap | ||
* submerge tubes in hot water bath for exactly 90s | * submerge tubes in hot water bath for exactly 90s | ||
* sit on ice for 5min | * sit on ice for 5min | ||
* add 1mL LB | * add 1mL LB | ||
| - | * | + | * incubate for 1h |
| - | * spread 500uL on Petri plate labelled with appropriate antibiotics | + | * spread 500uL on Petri plate labelled with appropriate antibiotics |
| - | * wait for | + | * Dip a glass rod with ethanol and flame it to spread cells in plates |
| - | * | + | * wait for plate to dry |
| + | * incubate overnight | ||
| - | [[University_of_Toronto_2006| Home]] | + | [[University_of_Toronto_2006| Home]] | [[Lab Training Protocol]] |
Latest revision as of 16:45, 28 June 2006
iGEM 2006 Lab Training Session I: Transformation
Key words: [http://en.wikipedia.org/wiki/Transformation_%28genetics%29 transformation], [http://en.wikipedia.org/wiki/Competent_cells competent cells]
Procedure
Preparation: competent cells
For each plasmid:
- take competent cells from -80C freezer
- thaw on ice for 10min
- add 2uL DNA to competent cell (label)
- mix with pipette
- sit on ice for 30min
- get a beaker of exactly 42C water from hot water tap
- submerge tubes in hot water bath for exactly 90s
- sit on ice for 5min
- add 1mL LB
- incubate for 1h
- spread 500uL on Petri plate labelled with appropriate antibiotics
- Dip a glass rod with ethanol and flame it to spread cells in plates
- wait for plate to dry
- incubate overnight
