Splitting Cells: Difference between revisions
Jump to navigation
Jump to search
changed serum names |
added temperature |
||
| Line 12: | Line 12: | ||
#Add 9 mL media to trypsinized cells | #Add 9 mL media to trypsinized cells | ||
#Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X) | #Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X) | ||
#Replace plates in the incubator | #Replace plates in the 37C incubator | ||
==Cell Specific Notes== | ==Cell Specific Notes== | ||
Revision as of 16:58, 10 February 2014
Materials
- Media (FBS or NCS as required): Prepare by filtering together 5 mL PSG, 50 mL COS-FBS and one bottle of DMEM
- PBS -/-
- 0.05% Trypsin
Protocol
- Warm PBS and Media in water bath
- Wash cells twice with 10 mL (per 10 cm dish) PBS -/-
- Add 1 mL trypsin and sit in the hood for 2-5 min
- Add 10 mL media to each new dish
- Check cells for trypsinization, and if necessary tap the cells
- Add 9 mL media to trypsinized cells
- Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X)
- Replace plates in the 37C incubator
Cell Specific Notes
- 3T3-L1 fibroblasts have special considerations regarding confluence. See Differentiation of 3T3-L1 Cells
- RAW 264.7 cells are scraped, not trypsinized. See Culturing RAW 264.7 Cells
- S2 cells are grown at 28C without extra CO2. See Culturing S2 Cells