Difference between revisions of "Freezing of C2C12 cells"
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(Created page with "Materials: - 15 ml FALCON tube - PBS w/o Ca2+ and Mg2+ - TE in PBS - Freezing media (10% DMSO + 40% serum + 50% DMEM) - Pipettes and tips - Glas pipetts - cryo tube - ice - f...") |
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Materials: | Materials: | ||
| − | + | * 15 ml FALCON tube | |
| − | + | * PBS w/o Ca2+ and Mg2+ | |
| − | + | * TE in PBS | |
| − | + | * Freezing media (10% DMSO + 40% serum + 50% DMEM) | |
| − | + | * Pipettes and tips | |
| − | + | * Glas pipetts | |
| − | + | * cryo tube | |
| − | + | * ice | |
| − | + | * freezing box | |
| − | |||
procedure: | procedure: | ||
Revision as of 07:33, 24 October 2014
Materials:
- 15 ml FALCON tube
- PBS w/o Ca2+ and Mg2+
- TE in PBS
- Freezing media (10% DMSO + 40% serum + 50% DMEM)
- Pipettes and tips
- Glas pipetts
- cryo tube
- ice
- freezing box
procedure:
- wash the cells with 5 ml PBS - remove the PBS and add 2 ml TE - let it incubate for 3 – 5 min - stop the TE with 5 ml HS – media and resuspend the cells - pipette the cell suspension back in the FALCON tube - centrifuge the cells (1000 xg, 3 min at 4 ºC) - remove the old media - resuspend the cells 1 ml freezing media - quickly pipette the liquid in the cryo tube - put the cells in the freezing box and store this for one day at –80 ºC - place the cells one day later into the liquid nitrogen