Splitting Cells

From Bridges Lab Protocols
Revision as of 15:08, 6 May 2009 by 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

  • 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

  1. Warm PBS and Media
  2. Wash cells twice with 10 mL (per 10 cm dish) PBS -/-
  3. Add 1 mL trypsin and sit in the hood
  4. Add 10 mL media to each new dish
  5. Check cells for trypsinization, and if necessary tap the cells
  6. Add 9 mL media to trypsinized cells
  7. Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X)
  8. Replace plates in the incubator