A sample protocol is listed here for transfection experiments performed in 6-well plates. To perform transfection experiments in other cell culture plates, simply multiply the suggested quantities by the relative surface area of your plate. GenScript recommends using Lipofectamine 2000 for all transfections. It consistently produces high transfection efficiency and high protein overexpression.
Add the 500 μl of DNA-Lipofectamine 2000 complexes to each well containing cells and medium. Mix gently by rocking the plate back and forth.
Incubate the cells at 37°C in a CO2 incubator for 24-72 hours until they are ready to assay for transgene expression. It is not necessary to remove the complexes or change the medium; however, growth medium may be replaced after 4-6 hours without loss of transfection activity.