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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?


Thanks, Aby

-abyasemani-

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

QUOTE (abyasemani @ Aug 29 2006, 11:34 AM)
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?


Thanks, Aby

-tertu-

QUOTE (abyasemani @ Aug 29 2006, 02:34 AM)
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?


Thanks, Aby


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?

-smartsunny-