I test out my electroporation effiency by using GFP plasmid, hence i figure out condition tat cause lesser cell death and reasonable tfx effiency (by eyes).

However, when i switch to my plasmid construct using exactly the same condition ie. electroporation medium, Volt, resistance), it caused a lot of cell death, hence the protein expression was low when i did the assaying.

The only diffences is tat my GFP is dissolved in dH2O while my construct is dissolved in TE buffer. Next, the concentration of both plasmid is not the same hence the volume added was vary.

could it be the reason above causing the problem or else there is other possiblities ??

thanks

I used to transfect lymphoma cells by elecroporation (in 0.5ml RPMI containing 10% FCS) and all our constructs were always in TE at 1µg/µl. Unless you we adding large volumes of your construct compared with the GFP I wouldn't have thought that having them in water or TE would make a difference.

Would you expect over-expression of your protein to be toxic or cause apoptosis? Maybe you could titrate the amount of plasmid down to see if you get reduced cell death or use an inducible expression vector to rule this out?

Ceri