transient transfection of a stable cell line? - (Nov/20/2006 )

hello everybody
i am trying to do a transient transfection in a stable 293A cell line but seems not working?
i am using lipofectamine for transfection....
any suggestion are mostly welcomed
thankxxxxxxxx

-spanishflower-

try to optimize transient transfection protocol with a reliable reporter vector such as pEGFP which bears the opportunity to monitor directly for fluorescence

-The Bearer-

in 293cells, the ratio 1µg DNA for 3µl lipofectamine showed good efficiency when transfected with pLN DsRED plasmid monitored under fluorescence microscope

-fred_33-

thankx guys...

i am using 4µg DNA and 10µl lipofectamine and its working in transient transfection of parent 293A cells so i was wondering why is it not as efficient for the stable cell line? i did a comparison using the same plasmid but for transient transfection in stable 293A and for transient transfection in parent 293A and the yield was not soo high in the first type????

thankx

-spanishflower-

QUOTE (spanishflower @ Nov 21 2006, 09:09 PM)

make sure you have roughly the same cell density for both cell types. It should be 70-90% confluent when you transfect 293.

-genehunter-1-