stable transfection of MDAMB231 and Malme3M cells. - (Dec/12/2005 )
Hi, I am doing stable transfection of MDA231 and Malme-3M cell lines. For both cell lines, I add G418 (500 ug/ml) to the medium for selection. For Malme-3M cells, I found all the cells (control) died in 5 days and the ones with transfection, I didn't see any clones after 8 days. Does this means the conc. of G418 is too high? But with MDAMB231 cells, only small amount of cells (control) died. I wonder if I can add more G418 now to incrase the selection strength. Anybody has experience with these two cell lines? Thanks a lot and Happy holidays :-)
I have no experiences with these cell lines. But in general you should do a pre-test to find optimal selection marker concentrations. It can be very different for different cell lines. Just test different concentrations of G418 on non-transfected cells for a few days/weeks and see which is the lowest concentration letal for these cells. Do not go much higher than this. It is not necessary.
Good luck and also happy holidays to you!