User:Karmella Haynes/Notebook/PcTF Genomics/2014/03/17

Pc-TF Genomics

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03/17/14 Mammalian cell culturing: Thawed KAH126 & KAH154 lines Cell culture: Attempt to thaw PcTF stable lines Request from Karsten Rippe, Heidelberg... U2OS Flp-in T-REx (parental cell line with no integration) U2OS Flp-in T-REx with hPCD-TF U2OS Flp-in T-REx with hPCD-RFP Notes: hPCD-TF = KAH126 hPCD-RFP = KAH154 First attempt at thawing KAH126 failed. Waited about 5 or so days to see growth. Try again, but wait longer, see if small sub-population is viable & forms colonies Procedure Quick-thaw cells Add 1 ml thawed cells to 5 ml 100% FBS Pellet Resuspend and grow in 4 mL 20% FBS, 1% P/S McCoy's 5A in T25 flasks Grow at 37°C. Check every 5 days.