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Team:UIUC-Illinois/Notebook/Protocols/Electrocompetence
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| - | <center><h1> | + | <center><h1>Making electrocompetent E.Coli cells</h1></center> <br/> |
| + | |||
| + | What you need: <br/><br/> | ||
| + | - 50mL subculture of E.coli cells<br/> | ||
| + | - 50mL conicial tubes<br/> | ||
| + | - Cold 10% glycerol solution <br/> | ||
| + | - Autoclaved 1.5mL centrifuge tubes<br/><br/> | ||
| + | Procedure:<br/><br/> | ||
| + | 1. Divide liquid cell subculture into 50mL conical tubes and centrifuge for 10 minute at 4000rpm <br/> | ||
| + | 2. Discard supernatant and add 25mL cold 10% glycerol <br/> | ||
| + | 3. Stir the mixture by mixing and centrifuge for an additional 10 minutes<br/> | ||
| + | 4. Repeat steps 2-3 two more times<br/> | ||
| + | 5. Discard the supernatant and resuspend the cells in 1mL 10% glycerol<br/> | ||
| + | 6. Transfer to 1.5mL centrifuge tubes and centrifuge for 2 minutes<br/> | ||
| + | 7. Discard the supernantant and resuspend the cells in 2 volumes (cell volumes) 10% glycerol<br/> | ||
| + | 8. Store at -80˚C<br/><br/> | ||
| + | |||
</div> | </div> | ||
Latest revision as of 15:31, 4 June 2012
