PolyFect Transfection Reagent
For fast and easy DNA transfection of standard cell lines
- Optimized transfection using cell-specific protocols
- Fast procedure and easy handling
- High cell viability, low cytotoxicity
Cat. No. / ID: 301105
PolyFect Transfection Reagent is based on activated dendrimer technology optimized for the transfection of DNA into COS-7, NIH/3T3, HeLa, 293, and CHO cells.
PolyFect Reagent consistently delivers high transfection efficiencies (see figures " High transfection efficiencies in CHO and 293 cells" and " PolyFect Reagent with HeLa cells"). In contrast to many liposomal reagents, PolyFect Reagent enables transfection in the presence of serum without lowering of transfection efficiency.
PolyFect Transfection Reagent is a solution of specifically designed activated-dendrimers. The reagent consists of dendrimer molecules of a defined spherical architecture with branches radiating from a central core (see figure " Activated dendrimer"). The branches terminate at charged amino groups, which can interact with negatively charged phosphate groups of nucleic acids. PolyFect Reagent assembles DNA into compact structures (see figure " PolyFect–DNA interaction"), that bind to the cell surface and are taken into the cell by nonspecific endocytosis. The reagent buffers the pH of the endosome, leading to pH inhibitition of endosomal nucleases, which ensures stability of PolyFect–DNA complexes.
PolyFect Transfection Reagent is provided as a ready-to-use solution. The transfection procedure (see flowchart " PolyFect 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. No post-transfection removal of complexes or medium change/addition is necessary, making the procedure fast and easy.
Transfection using PolyFect Reagent requires minimal handling and removal of transfection complexes is not required, making this reagent highly suitable for high-throughput screening. PolyFect Reagent provides outstanding transfection efficiencies, excellent reproducibility, and minimal cytotoxicity in high-throughput 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.
PolyFect Transfection Reagent is highly suited for efficient transfection of standard cell lines in:
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.
|applications||Plasmid transfection, protein overexpression|
|transfectiontype||Transient transfection, stable transfection|
|features||Optimized protocols for COS-7, NIH/3T3, HeLa 293, and CHO cells. Transfection in the presence of serum|
|numberofpossibletransfections||Up to 50–100 transfections in 6-well plates / 1 ml reagent|
|celltype||COS-7, NIH/3T3, HeLa, 293, CHO cells|
QIAGEN Transfection Reagents have been used successfully with many different cell types. For your convenience, we have organized data from researchers who have shared their experimental results with us online in our Transfection Cell Database. Simply type your cell line of interest into the 'Search' field on this page, and find transfection results achieved with various QIAGEN Transfection Reagents. Please note that QIAGEN cannot verify data supplied from outside sources.
You can also submit your own transfection data and obtain a gift as appreciation. In addition you can find on our TransFect Protocol Database transfection protocols for specific cell types and plate formats.
For the transfection of oligonucleotides and/or large vector constructs (>20 kb) using PolyFect Transfection Reagent, we recommend performing optimization trials to determine the optimal Transfection Reagent/DNA ratio. The amounts of DNA and Reagent provided in the PolyFect Transfection Reagent Handbook can be used as a guideline.
We suggest to start optimization using a range between half and twice the recommended amount of PolyFect Transfection Reagent while keeping the DNA amount constant, as well as using half and twice the recommended amount of DNA while keeping the amount of PolyFect Transfection Reagent constant. For increasing the transfection efficiency of adherent cells you can use the Attractene Transfection Reagent, or of primary cells and difficult-to-transfect cells using Effectene Transfection Reagent. "
Yes, please follow the Supplementary Protocol 'Transient Transfecton of HeLa-S3 cells in 60 mm dishes using PolyFect Transfection Reagent' (TFP01).
Yes, we have Supplementary Protocols for different cell lines:
The composition of 1x PBS solution is:
Adjust to a final pH of 7.4.
This solution is not supplied in any QIAGEN Kit, but is used in protocols for various QIAGEN transfection kits.
Yes, we have a Supplementary Protocol for 60 mm dishes and 96-well plates: