PolyFect Transfection Reagent is provided as a ready-to-use solution. The transfection procedure is fast and simple just add PolyFect Reagent to the DNA solution, mix, incubate for 510 minutes, add growth medium (which can contain serum and antibiotics), and pipet the PolyFectDNA complexes onto the cells. The cells are then incubated for gene expression. No post-transfection removal of complexes or medium change/addition is necessary, making the procedure fast and easy.
PolyFect Reagent has been specifically developed for fast and easy transfection of COS-7, NIH/3T3, HeLa, 293, and CHO cells. Optimized, cell-specific protocols for transfection of these cell lines in different cell-culture formats are provided in the PolyFect Transfection Reagent Handbook. In addition, optimized protocols for transfection of HeLa-S3 cells are available from QIAGEN Technical Services and the Transfection Tools web site (www.qiagen.com/transfectiontools/).
PolyFect Reagent consistently delivers high transfection efficiencies
(see figures "High Transfection Efficiencies"
and "PolyFect Reagent with HeLa Cells").
ntrast to many liposomal reagents, PolyFect Reagent enables transfection
in the presence of serum without lowering of transfection efficiency.
High Transfection Efficiencies
Comparison of transfection efficiencies for PolyFect Reagent and two lipid-based reagents. Cells cultured in 6-well plates were transfected with a beta-galactosidasereporter 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 l Reagent L2 with 3 g DNA and 15 l Reagent F with 2 g DNA for CHO cells, and 10 l Reagent L2 with 2 g DNA and 7 l Reagent F with 3 g DNA for 293 cells. All transfections were performed in triplicate. Two independent transfection experiments are shown for 293 cells.
PolyFect Reagent with HeLa Cells
Expression of A -galactosidase and B green fluorescent protein (GFP) in HeLa cells. Cells were cotransfected in 6-well plates with -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.
The application of recombinant DNA technology to fields such as drug discovery and development has led to an increased need for high-throughput transfection. Transfection using PolyFect Reagent requires minimal handling and removal of transfection complexes is not required, making this reag ent highly suitable for high-throughput screening. PolyFect Reagent provides outstanding transfection efficiencies, excellent reproducibility, and minimal cytotoxicity in highthroughput transfection, and is available in bulk quantities. For optimized protocols for transfection of COS-7, NIH/3T3, HeLa, HeLa-S3, 293, and CHO cells in 96-well plates, please contact QIAGEN Technical Services or visit the Transfection Tools web site at www.qiagen.com/transfectiontools/.
1. Tang, M.X., Redemann, C.T., and Szoka, Jr., F.C. (1996) In vitro gene delivery by degraded polyamidoamine dendrimers. Bioconjugate Chem. 7, 703.