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Team:Glasgow/Notebook/Protocols
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| - | + | <h2>Creating Chemically Competent Cells</h2> | |
<ol> | <ol> | ||
| - | <li></li> | + | <li>Pipette 400μl of culture into 20ml of fresh broth</li> |
| - | <li></li> | + | <li>Incubate the cultures in shaking 37⁰C water bath for 90 min</li> |
| - | <li></li> | + | <li>Calcium chloride and centrifuge tubes must be cooled on ice.</li> |
| - | <li></li> | + | <li>Pour culture into centrifuge tubes, return to ice.</li> |
| - | <li></li> | + | <li>Centrifuge in a cold rotor for 2 min at 7000rpm (6000 G) </li> |
| - | <li></li> | + | <li>Pour out the supernatant, being careful about the pellet</li> |
| - | <li></li> | + | <li>Add 10μl of calcium chloride, immediately return to ice and leave for 1 hour (or longer)</li> |
| - | <li></li> | + | <li>Repeat the centrifuge step, pour out supernatant</li> |
| + | <li>Store on ice, add 1ml of calcium chloride. Resuspend and return to ice.</li> | ||
</ol> | </ol> | ||
Revision as of 14:42, 15 October 2014
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Protocols
Edit This page shows our protocols for various lab procedures.
Creating Chemically Competent Cells
- Pipette 400μl of culture into 20ml of fresh broth
- Incubate the cultures in shaking 37⁰C water bath for 90 min
- Calcium chloride and centrifuge tubes must be cooled on ice.
- Pour culture into centrifuge tubes, return to ice.
- Centrifuge in a cold rotor for 2 min at 7000rpm (6000 G)
- Pour out the supernatant, being careful about the pellet
- Add 10μl of calcium chloride, immediately return to ice and leave for 1 hour (or longer)
- Repeat the centrifuge step, pour out supernatant
- Store on ice, add 1ml of calcium chloride. Resuspend and return to ice.
