Splitting Cells: Difference between revisions
| Line 7: | Line 7: | ||
#Warm PBS and Media in water bath | #Warm PBS and Media in water bath | ||
#Wash cells twice with 10 mL (per 10 cm dish) PBS -/- | #Wash cells twice with 10 mL (per 10 cm dish) PBS -/- | ||
#Add 1 mL trypsin and sit in the hood | #Add 1 mL trypsin and sit in the hood for 2-5 min | ||
#Add 10 mL media to each new dish | #Add 10 mL media to each new dish | ||
#Check cells for trypsinization, and if necessary tap the cells | #Check cells for trypsinization, and if necessary tap the cells | ||
| Line 13: | Line 13: | ||
#Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X) | #Add 1 mL cells to each dish (for 10X dilution; 0.5 mL for 20X) | ||
#Replace plates in the incubator | #Replace plates in the incubator | ||
==Cell Specific Notes== | ==Cell Specific Notes== | ||
*3T3-L1 fibroblasts have special considerations regarding confluence. See [[Differentiation of 3T3-L1 Cells]] | *3T3-L1 fibroblasts have special considerations regarding confluence. See [[Differentiation of 3T3-L1 Cells]] | ||