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.
Binod G C
I'm Binod G C (MSc), a PhD candidate in cell and molecular biology who works as a biology educator and enjoys scientific blogging. My proclivity for blogging is intended to make notes and study materials more accessible to students.
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