selecting the transfected colonies after transfection - (Aug/28/2006 )
hi everybody. before transfection, i am using different concentrations of hygromycin first on my host cells to determine the best dose for selection. my cells are Chinese hamster cell lines. in the selection prptocols they say after adding the media plus antibiotics to the media you have to change the media every 2-3 days. i start with very low seeding density. for 3 days it is fine and no need to change the media but then the meida turn yellow and overpoulated very fast. in less than 24 hours after changing the media. what should i do? considering that we have to allow about 3 weeks for selecting the transfected cells.
thanks,
Aby
if it's fine as u said, u don't need to change the media each 2-3 days, u can change it after more few days
u can modify your protocol, don't stuck to every word written
that's what i learnt