I am doing transfection for luciferase assay. How can I determine the amount of plasmid encoding gene of interest and plasmid encoding luciferase? Many thanks!
I'm not really sure what your question is. How do you want to to optimize the ratio? What are you looking for, maximum luciferase expression? Minimum amount of luciferase plasmid you can use and still detect it?
The standard way to optimize this type of thing is to add the two plasmids at different ratios (gene of interest:luciferase) of 1:1, 2:1, 5:1, 10:1, ....etc. and then measure luciferase and gene of interest expression and see what condition is best for your purposes.
Luciferase is generally one of the most sensitive reporter genes, so it is likely you will be able to get away with a very low amount (10:1 or lower) of luciferase plasmid and still have enough to reliably detect by luciferase assay.
Good luck....
Thank you Jonathan. That's what I want. Since it is quite sensitive, then I will start with a low amount of luciferase plasmids (gene of interest: luciferase = 5:1).
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