I am PhD student and I did a transfection experiment for three tumour cells lines. I found all my cells were dead. Please help me.My experiment was as below.
I tried to optimize the transfection with turbofectin 8 (origene) and lipofectamin (Invitrogen) for three cell lines. I did according to the manufactures guidelines. Cells were live and were in a more than 90% confluence before I tripsynized them after 48 hrs for FACS analysis. Cells were in EMEM and DMEM (Complete medium). As I was optimizing the protocol, used the pmax-GFP plasmid and I washed the cells in PBS. I centrifuged the cells to remove PBS and found cells in clumps. For FACS, resuspended the cells in PBS and didn't do cell fixation.
Please help me.