The transfection procedure is fast and simple - just add PolyFect Reagent to the DNA solution, mix, incubate for 5-10 minutes, add growth medium (which can contain serum and antibiotics), and pipet the PolyFect-DNA complexes onto the cells. The cells are then incubated for gene expression.
Expression of [A] beta-galactosidase and [B] green fluorescent protein (GFP) in HeLa cells is shown. Cells were cotransfected in 6-well plates with beta-galactosidase- and GFP-reporter plasmids using PolyFect Transfection Reagent and the HeLa cell protocol. Expression was visualized by X-gal staining or fluorescence microscopy 2 days post-transfection.
Transfection efficiencies for PolyFect Reagent and 2 lipid-based reagents were compared. Cells cultured in 6-well plates were transfected with a beta-galactosidase-reporter plasmid. The appropriate protocol was used for transfection with PolyFect Reagent. Optimized protocols were used for transfection with Reagents L2 and F; these were: 10 µl Reagent L2 with 3 µg DNA and 15 µl Reagent F with 2 µg DNA for CHO cells, and 10 µl Reagent L2 with 2 µg DNA and 7 µl Reagent F with 3 µg DNA for 293 cells. All transfections were performed in triplicate. Two independent transfection experiments are shown for 293 cells.