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reproducibility of luciferase assay - (Oct/24/2008 )

First of all, I am a chemistry guy and I worked on transfection for half year. I faced problem in reproducibility. (well, I faced it in MTT assay too) I hope someone can help me. THX!
I synthesized some cationic polymers and wanna to test for its transfection efficiency and I used luciferase assay to quantify it. The RLU are always different a lot, even with transfection kit (+ve control).
I use 96 well plate with 5x10^3 cell/well and look for 48 hr expression, triplicate, reagents were mixed 3.5 times and 1 portion added to wells. I worked on 2 96 well plates in a time, cell lines were U87 and U373.

reply if anyone can help!
Thank you!!!

-leosiu-

You will find something interesting in JALA (Journal of Association for Laboratory Automation)

some years ago they issued two good tutorials:

1) Assay Optimization: A Statistical Design of Experiments Approach (doi:10.1016/j.jala.2005.11.001)
2) Efficient Assay Development and Screening at a Small Research Site (doi:10.1016/j.jala.2006.02.001)

-96well-