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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...'
==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
*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
