Difference between revisions of "Team:KU Leuven/Experiments"
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| − | <h3> | + | <h3>Wet Lab and bacterial lab</h3> |
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| − | <h3> | + | <h3>Cell Culture</h3> |
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| − | + | <b>Medium of HEK T293 cells</b> | |
| − | <br/> | + | <br> |
| + | <ul> | ||
| + | <li>DMEM (Gibco N° 41965-039) 500 ml</li> | ||
| + | <li>50 ml (10 %) FCS = (Hyclone)</li> | ||
| + | <li>10 ml Penicillin/Streptomycin (Gibco N° 15070-071) = 100-100U/ml</li> | ||
| + | <li>10 ml Glutamax (Gibco N° 35050-038) = 10 mM</li> | ||
| + | <li> 5 ml Non-Essential-Amino-Acids (Gibco N° 1140-035) = 10x</li> | ||
| + | </ul> | ||
| + | <br> | ||
| + | <br> | ||
| + | <b>Growing Conditions</b> | ||
| + | <br> | ||
| + | <ul> | ||
| + | <li>Incubate at 37°C and 10% CO2</li> | ||
| + | <li>Split cells around 70-90% confluency</li> | ||
| + | <li>Change medium twice a week</li> | ||
| + | </ul> | ||
| + | <br> | ||
| + | <br> | ||
| + | <b>Standard procedure for Trypsinization</b> | ||
| + | <br> | ||
| + | <ol type="1"> | ||
| + | <li>Wash the cells with Versene (Gibco N° 15040-033, ± 2 min)</li> | ||
| + | <li>Remove versene and add Trypsin (Gibco N° 25300-096)</li> | ||
| + | <li>Incubate at 37°C (around 3-5 min)</li> | ||
| + | <li>Stop the trypsinization by adding 5-10 times volume of medium</li> | ||
| + | <li>Count cells</li> | ||
| + | <li>Centrifuge for 5 minutes at 1200 rpm</li> | ||
| + | <li>Aspirate supernatant and resuspend the cells in medium</li> | ||
| + | <li>Plate cells for experiments or culture</li> | ||
| + | </ol> | ||
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| + | <br> | ||
| + | <b>Coating | ||
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| − | <h3> | + | <h3>Electrophysiology Lab</h3> |
</div> | </div> | ||
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Latest revision as of 13:54, 30 August 2017
Experiments
In the lab, we used different experimental procedures. There are protocols for the wet and bacterial lab, for the cell culture lab and for the electrophysiology lab.
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