stable transfection for suspension cell lines - (Jun/25/2007 )

Dear all,

I would like to transfect Jurkat cell lines. These cells are in suspension and I would like to do some stable transfection on it. The first think is to find the rigth concentration of blasticidin for the transfection. Does someone has a protocol for that? I know it is easy to determine death cells when we transfect adherent cells but how could I do with cells in suspension?

Vanessa Suin

Can you do : 1) left the cells along for several days and see medium color change/cell growth, or 2) do a cell counting after trypan blue staining under light microscope after several days. You have to wash cells once. Dead cells will be stained blue. PI staining for nuclei/fluorescence microscopy works as well. If you use just the right amount of PI, such that the background is not to high, you dont have to wash the cells. Dead cells will be stained bright red.