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Team:EPF-Lausanne/Notebook/19 July 2012
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| - | '''Cell counting''' | + | ; '''Cell counting''' |
Given our cell density we used a 12x dilution (table). We transfered 6 samples of cells into a 96 well plate, added Trypan blue and analysed each sample with Countess. | Given our cell density we used a 12x dilution (table). We transfered 6 samples of cells into a 96 well plate, added Trypan blue and analysed each sample with Countess. | ||
[table with results] | [table with results] | ||
| - | '''Western Blot sampling''' | + | ; '''Western Blot sampling''' |
Protocol (add to separate page): | Protocol (add to separate page): | ||
Revision as of 21:48, 19 July 2012
Contents |
Morning: Cell counting and sampling
- Cell counting
Given our cell density we used a 12x dilution (table). We transfered 6 samples of cells into a 96 well plate, added Trypan blue and analysed each sample with Countess.
[table with results]
- Western Blot sampling
Protocol (add to separate page):
- Centrifugation of the cell samples at 2500xg for 10min. Discard the supernatant (vacuum).
- Washing: Resuspend the pellet in PBS1x, centrifuge at 2500xg for 10min, discard supernatant.
- Add at least 1mL PER-M Reagent for 100mg (~100µl) of cell pellet. Pipette up and down to suspend.
- Shake gently for 10min.
- Remove cell bedris by centrifugation: ~1400xg for 15min
- Transfer the supernatant into new tubes for analysis or storage.
- Samples where taken from odd numbered tubes : 1,3,5,7.
- We used 300µl of M-PER Reagent based on the size of our pellets.
- Maximum centrifugation was 10'600xg.
Protocol: None
Forgot to insert protocol.
- Comments
For samples 3 and 5 we had to repeat step 5 (centrifugation to remove cell debris) because the cell pellet got aspired in the pipette while trying to transfer the supernantant into fresh tubes.
Afternoon: Cell lysates
Protocol: None
Forgot to insert protocol.
- Comments
