- Open the cell culture hood, turn on the light and the blower. Make sure to clean the hood and wipe it with 70% ethanol before use..
- Vacuum out the media from the cell culture plate.
- Add PBS (1-2 ml) to wash the plate. Add PBS from the wall of the plate and mix it by rotating the plate.
- Pipette out the PBS using pipette aid
- Add Trypsin (1ml). It can be added right on the top of the cell. Spread well.
- Incubate in CO2 incubator for 10 minutes. Time may vary depending on weather the cells are not adherent anymore or not.
- 1-2 ml of culture medium can be added and mixed well by pipetting to make sure no clumps are present.
- Discard the medium, keeping around 0.3 – 0.5 ml of the medium in the plate.
- Add the medium to make 8-10 ml to dilute and rotate it to spread the cells well.
- Incubate in CO2 incubator.
