1 reply to this topic

[flyingbullet]

Hey all, I need help

I transfected SH-SY5Y cells with a protein in a pCDNA vector and selected them using geneticin 250µg/ml.
1) the untransfected control plate died 1-2 days later - on the transfected plates only about 80 % died and the rest looked well.
2) the transfected cell do not grow. they are 2 weeks under selection now and seem to be fewer from day to day.
3) the untransfected cells that are not under selection still grow well, so its not a problem of the medium (AMEM, F12, FBS, Pen/Strep, L-Glu)

Where could the problem be???

[bob1]

What density are the cells at?  Selection only works when the cells are actively growing, this means that you typically need them at less than 70% confluency.