User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/08/04
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Expand the Thawed cells
Cells became 100% confluent in T-25 flask. wash the cells with 2 mL 1xPBS, trypsinize them with 0.5 mL of Trypsin buffer and transfer the entire culture into a T-75 flask with 10 mL total growth medium (10% FBS, 1% pen-strep). After the t-75 is 100% confluent, you can make new frozen stocks or passage the cells at 1:10 (or 1:5 if they are slow-growing).