I did lipofectamine transfection in 12-well plates with 0.5M of cells and 0.8ug of TOPFLASH plasmid in each well. After 24h, I changed to media to serum-free media supplemented with 10uM CHIR99021 or 50ng/uL of Wnt3a and measure the luciferase signal the next day. However, I have trouble getting consistent results. Sometimes I able to see induction by CHIR in 18h - 24h but sometimes I don't, can anyone please give me some advice?
Junjie Shao
We always add a beta-gal expressing plasmid as transfection efficiency marker.
and then use the fluc/beta-actin as result. Rluc works the same way.
if you do not included Rluc, your transfection need to be very consistent.
sometimes, I found if the transfection efficiency is low, then you will see a high variation.
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