Difference between revisions of "Generating DMSO Stocks for Cell Culture"

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(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...')
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Revision as of 15:35, 13 August 2009

Materials

  • Cells in 10cm dishes, at 90-95% confluence.
  • Cryopreservation Container (Nalgene 5100-0001)
  • Sterile DMSO

Protocol

  1. Pick a low passage number of cells and grow 2-5 10cm dishes.
  2. At near confluence wash cells twice with PBS -/- and trypsinize normally
  3. Collect all the cells in a 15 mL falcon tube
  4. Centrifuge 5 min at 1500RPM to pellet cells
  5. Aspirate media
  6. Add media (1.8 mL per original plate)
  7. Add sterile DMSO to a final concentration of 10% (0.2 mL per original plate)
  8. Gently resuspend cells and aliquot 1 mL of suspension into cryopreservation vials
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