TC1-Alpha cells transfection redo
- TC1-Alpha cells transfected 10 days ago were dead due to overgrowth. Transfection needs to be redone.
- Two T-75 flasks(10mL at 100% confluence) of the TC1-Alpha Cells
Transfection (6 well)
- T-75(1) container used
- Each well done 1/10, 1mL detached cells to 3mL DMEM.
- 6 wells= 6/10 of one T-75 used.
- Rest of T75(1) container used.
- Split cells 1/5, 2mL detached cells to 8mL DMEM.
- Rest of T75(1) container disposed
- Protocol used for these two functions was the Splitting Cell protocol
- Concentration: 90% FBS, 10% DMSO
- T75(2) flask used, detach cells according to Splitting Cell protocol
- Transfer cells to spin column and spin down
- Aspirate fluid, leaving enough to cover top of pellet, re-suspend pellet
- Add 5mL Freezing Serum
- Remix cells in Freezing serum
- 1mL into tubes, 5 tubes created for freezing