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PolyFect Transfection Reagent

For fast and easy DNA transfection of standard cell lines

Features

  • Optimized transfection using cell-specific protocols
  • Fast procedure and easy handling
  • High cell viability, low cytotoxicity
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PolyFect Transfection Reagent (1 ml)

Cat. No. / ID: 301105

For 25–65 transfections in 60 mm dishes or 50–100 transfections in 6-well plates.
Quantity
1 ml
4 x 1ml
100 ml
The PolyFect Transfection Reagent is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

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.

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.

See figures

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.

See figures

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.

See figures

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

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPlasmid transfection, protein overexpression
Transfection typeTransient transfection, stable transfection
ControlsNot included
FeaturesOptimized protocols for COS-7, NIH/3T3, HeLa 293, and CHO cells. Transfection in the presence of serum
TechnologyActivated dendrimers
Number of possible transfectionsUp to 50–100 transfections in 6-well plates / 1 ml reagent
Nucleic acidDNA
Cell typeCOS-7, NIH/3T3, HeLa, 293, CHO cells

Resources

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.
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.
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
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.
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.
The following protocol is optimized for transient transfection of 293 cells in 96-well plates.
The following protocol is optimized for transient transfection of CHO cells in 96-well plates.
The following protocol is optimized for transient transfection of COS-7 cells in 96-well plates.
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.
The following protocol is optimized for transient transfection of NIH/3T3 cells in 96-well plates.
Brochures & Guides (1)
Brochure detailing reagents for efficient and robust DNA and RNA transfection.
Kit Handbooks (1)
For optimized transfection of COS-7, NIH/3T3, HeLa, 293, and CHO cells
Quick-Start Protocols (1)
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.

FAQ

What are your recommendations for cotransfecting several plasmids?
The recommended amount of DNA being transfected should be split between the different plasmids. For example, if you normally transfect 5 µg of one plasmid, use 2.5 µg each for two plasmids (other proportions of the two plasmids can be used, as long as 5 µg total is maintained) as a starting point for optimization.
FAQ ID -124
Do you have transfection data for QIAGEN Transfection Reagents?

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.

FAQ ID -158
How can I optimize the transfection of oligos and large plasmids using PolyFect Transfection Reagent?

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. "

FAQ ID -176
Do you have a protocol for transient transfection of HEK 293 cells in 75 cm3 flasks?
FAQ ID -1014
Do you have a protocol for transient transfection of HeLa-S3 cells in 60 mm dishes using PolyFect?

Yes, please follow the Supplementary Protocol 'Transient Transfecton of HeLa-S3 cells in 60 mm dishes using PolyFect Transfection Reagent' (TFP01).

FAQ ID -1012
Do you have a protocol for transient transfection of cells in 96-well plates using PolyFect Transfection Reagent?
What is the recipe for 1x PBS solution?

The composition of 1x PBS solution is:

  • 137 mM NaCl
  • 2.7 mM KCl
  • 4.3 mM Na2HPO4
  • 1.47 mM KH2PO4

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.

FAQ ID -1030
Do you have a protocol for transient transfection of Hela-S3 cells using PolyFect Transfection Reagent?

 Yes, we have a Supplementary Protocol for 60 mm dishes and 96-well plates:

FAQ ID -997