Lipofectamine for Transfecting Plasmid DNA in Cell Lines and Primary Hippocampal Neurons

We use Lipofectamine 2000 to transfect plasmid DNA to silence our protein of interest in human colorectal cancer cell line Caco2, HEK, primary hippocampal neurons and also ependymal cells. Lipid DNA complex is made by diluting DNA and lipofectamine separately in optiMEM media and mixing them in the ratio of 1:1 after 15-20 mins of incubation. The mixture will be then added to the cells and the media is changed after 2 hours to complete the transfection. High transfection efficiency is achieved with Lipofectamine 2000 reagent and it can be used in serum supplemented media also.