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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

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.

Cat No./ID: 301105
PolyFect Transfection Reagent (1 ml)
$183.00
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For 25–65 transfections in 60 mm dishes or 50–100 transfections in 6-well plates.
Cat No./ID: 301107
PolyFect Transfection Reagent (4 x 1 ml)
$657.00
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For 100–260 transfections in 60 mm dishes or 200–400 transfections in 6-well plates.
Cat No./ID: 301108
PolyFect Transfection Reagent (100 ml)
$13,582.00
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For 2500–6500 transfections in 60 mm dishes or 5000–10,000 transfections in 6-well plates.
The PolyFect Transfection Reagent is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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High transfection efficiencies in CHO and 293 cells.

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.

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PolyFect transfection procedure.
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.
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Activated-dendrimer structure.

Note the highly branched structure of an activated dendrimer in this schematic representation.

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PolyFect–DNA interaction.

A model of the PolyFect-DNA complex shows PolyFect Reagent (purple balls) interacting with DNA (black) to form a ring-like (toroid-like) structure. The upper right section of the illustration shows naked DNA, the lower section shows the interaction between dendrimers and DNA inside the complex, and the upper left section shows the final complete coverage of DNA within the complex.

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PolyFect Reagent with HeLa cells.
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.
Performance

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.

Principle

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.

Procedure

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.

High-throughput transfection

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.

Applications

PolyFect Transfection Reagent is highly suited for efficient transfection of standard cell lines in:

Studies on gene expression and function
Drug discovery and development studies
Features
Specifications
Applications Plasmid transfection, protein overexpression
Cell type COS-7, NIH/3T3, HeLa, 293, CHO cells
Controls Not included
Features Optimized protocols for COS-7, NIH/3T3, HeLa 293, and CHO cells. Transfection in the presence of serum
Nucleic acid DNA
Number of possible transfections Up to 50–100 transfections in 6-well plates / 1 ml reagent
Technology Activated dendrimers
Transfection type Transient transfection, stable transfection

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Brochures & Guides (1)
Brochure detailing reagents for efficient and robust DNA and RNA transfection.
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Kit Handbooks (2)
For optimized transfection of COS-7, NIH/3T3, HeLa, 293, and CHO cells
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COS-7、NIH/3T3、HeLa、293、CHO細胞に至適化したトランスフェクション用
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Transfection Protocols (2)
Search for transfection data by nucleic acid, cell line, and transfection reagent. Our database contains data from researchers like yourself who have shared their experimental results with us.
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Transfection protocols for specific cell types and plate formats that save you the time and effort of adapting existing protocols to fit your requirements. Simply select the cell type, nucleic acid, and culture format to receive a QIAGEN transfection protocol to print out or download in convenient PDF format.
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Supplementary Protocols (13)
The following protocol is optimized for transient transfection of HeLa-S3 cells in 60 mm dishes. Parameters for transfection using other culture formats are given in Table 1. Please note that this protocol is only for HeLa-S3 cells. Optimized protocols for HeLa cells are provided in the PolyFect® Transfection Reagent Handbook.
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The following protocol is optimized for transient transfection of HeLa-S3 cells in 96-well plates. Please note that for HeLa cells the optimal amounts of DNA and PolyFect® Reagent differ from the amounts given in the protocol below. Please call one of the QIAGEN Technical Service Departments or your local distributor for an optimized HeLa protocol.
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The following protocol is optimized for transient transfection of 293 cells in 96-well plates.
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The following protocol is optimized for transient transfection of CHO cells in 96-well plates.
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The following protocol is optimized for transient transfection of COS-7 cells in 96-well plates.
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The following protocol is optimized for transient transfection of HeLa cells in 96-well plates. Please note that for HeLa-S3 cells the optimal amounts of DNA and PolyFect® Reagent differ from the amounts given in the protocol below. Please call one of the QIAGEN Technical Service Departments or your local distributor for an optimized HeLa-S3 protocol.
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The following protocol is optimized for transient transfection of NIH/3T3 cells in 96-well plates.
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User-Developed Protocols (1)
This procedure has been developed by customers for the transient transfection of HEK 293 cells in 75 cm3 flasks using QIAGEN® PolyFect® Transfection Reagent. It has not been thoroughly tested and optimized by QIAGEN.
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