01/06/2006 GS
Splitting Cells
1. Warm Regular HT22, HT22 w/Hygromycin (if needed), and Trypsin/EDTA (T/E)
media for 30 min in water bath
2. Wash bottle off with ethanol
3. Take flasks of cells from incubator turn them upright and suck off all old media with
glass pipette.
4. Add 5 ml of Tryp EDTA
5. Place flasks back into incubator for 3 min
6. After 3 min, look at flasks under microscope to make sure cells are floating
7. Add 5 ml of fresh media into the flask, and swish around gently (make sure that when
adding media, that there are at least equal amounts of media to Trypsin)
8. Take 2 new flasks and label with date, passage number, type of cells and split ratio.
(One is a back flask)
9. Add 8 ml of fresh media to the new flasks
10. Add 2 ml of media/Trypsin from old flasks to each of the new flasks and mix gently.
This is a 1:5 split.
11. Place new flasks in incubator.