Difference between revisions of "Splitting Cells"
From Bridges Lab Protocols
Davebridges (Talk | contribs) (Created page with '==Materials== *FBS (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== #W...') |
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==Materials== | ==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 -/- | *PBS -/- | ||
*0.05% Trypsin | *0.05% Trypsin | ||
==Protocol== | ==Protocol== | ||
| − | #Warm PBS and Media | + | #Warm PBS and Media in water bath |
#Wash cells twice with 10 mL (per 10 cm dish) PBS -/- | #Wash cells twice with 10 mL (per 10 cm dish) PBS -/- | ||
#Add 1 mL trypsin and sit in the hood | #Add 1 mL trypsin and sit in the hood | ||
Revision as of 15:30, 6 May 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
