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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...'
==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==
#Warm PBS and Media
#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
*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==
#Warm PBS and Media
#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
