HELP! About plasmid transfection in 96 well plate - (Jun/27/2007 )

I was doing some experiment which need transfect reporter vector in 96 well plate. I had some experience in 24-well transfection, so I minimized the transfection system to 1/5 of original one. However, when I assay the luminensence (I use Renilla luciferase), the result (RLuc activity) in 96 well plate is far less than the result in 24 well plate. Why does this happen? It is supposed to be the same result 'cause every condition is minimized proportionally.

Could anybody give me an advice? Thanks

I found out that you need to use more DNA for 96-wells (0.2 to 0.4 ug per well), while keep the DNA to transfection reagent ratio the same. Try it and tell us what happens.


Be sure not to use cells that are confluent, that is important.

Thank you, I'll try it soon~~

Sorry I meant to say NOT confluent.