Luciferase concentration question - (Oct/16/2007 )

Hello everyone,

I have a quick question....I am doing a luciferase assay using 2 plasmids. I want to keep the concentration of 1 of the plasmids constant (500ng) and vary the concentration of the other plasmid (0, 10, 50, 100, 250 and 500ng). How can I do that? From what I understand, during transfection with lipofectamine, I would need to keep the concentration constant and equal between different wells! I would appreciate your insights and experience.

Thanks in advance,
Una

You need to use a third, unrelavent plasmid, such as an empty expression vector, as the stuff or filling plasmid to bring up the total amount of plasmid to the same for all groups.

Hi,

genehunter-1 is right. You can use pUC18 or pUC19 for instance (500, 490, 450, 400, 250, 0ng).

Thanks genehunter-1 & zek....I will do that.

Thanks,
Micron