Generating DMSO Stocks for Cell Culture
From Bridges Lab Protocols
Revision as of 15:35, 13 August 2009 by Davebridges (Talk | contribs) (Created page with '==Materials== *Cells in 10cm dishes, at 90-95% confluence. *Cryopreservation Container (Nalgene 5100-0001) *Sterile DMSO ==Protocol== #Pick a low passage number of cells and gro...')
Materials
- Cells in 10cm dishes, at 90-95% confluence.
- Cryopreservation Container (Nalgene 5100-0001)
- Sterile DMSO
Protocol
- Pick a low passage number of cells and grow 2-5 10cm dishes.
- At near confluence wash cells twice with PBS -/- and trypsinize normally
- Collect all the cells in a 15 mL falcon tube
- Centrifuge 5 min at 1500RPM to pellet cells
- Aspirate media
- Add media (1.8 mL per original plate)
- Add sterile DMSO to a final concentration of 10% (0.2 mL per original plate)
- Gently resuspend cells and aliquot 1 mL of suspension into cryopreservation vials
