Difference between revisions of "Splitting Cells"
From Bridges Lab Protocols
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#Replace plates in the incubator | #Replace plates in the incubator | ||
==Cell Specific Notes== | ==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]] | *RAW 264.7 cells are scraped, not trypsinized. See [[Culturing RAW 264.7 Cells]] | ||
Revision as of 19:15, 30 June 2009
Materials
- Media (L1-FBS for 3T3-L1, COS-FBS for others): 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
- 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 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
