Getting rid of the dead cells during selection of transfected colonies? - (Aug/29/2006 )
In most of the protocols for selection of the transfected cells we have to change the media plus antibiotics every 2-3 days. However, my dead cells seem to attach very strongly to the media and occupy all the space and make the condition bad for the surviving cells. Therfore meida turns yellow very fast . how can i get rid of the dead cells and keep a good condition for resistant cells during 2-3 weeks of selection?
how do u know they are dead? if itturns yellow, that means theres a lot of consumption.. maybe from the cells you think they are dead.
Neomycin selection can last 3 or 4 weeks!
with puro 3 days to 5
I have been told that dead cells float after detaching from the plate. Are u sure ur cells are dead. Y don't u remove the so-called "dead" cells & do a trypan blu exclusion test? My cells float when the population density is too high or if they are dead. And ur media is turining yellow which means there ia a lot of metabolic activity in ur plates. There isn't any bacterialc ocntamination, is there?