PolyFect Transfection Reagent (1ml)

A DNA transfection reagent from QIAGEN built on activated dendrimer technology for fast delivery of plasmid DNA into standard mammalian cell lines. The reagent is a ready-to-use solution of specifically designed activated-dendrimers with a defined spherical architecture whose branches radiate from a central core and terminate at charged amino groups. These positively charged termini interact with negatively charged phosphate groups on nucleic acids, assembling DNA into compact structures that bind to the cell surface and enter cells via nonspecific endocytosis. Once internalised, the reagent buffers the pH of the endosome, inhibiting endosomal nucleases and helping to preserve the integrity of the reagent–DNA complexes.

Optimised protocols are available for COS-7, NIH/3T3, HeLa, 293 and CHO cells, supporting plasmid transfection and protein overexpression in both transient and stable formats. In contrast to many liposomal reagents, transfection can be performed in the presence of serum without a reduction in transfection efficiency. The workflow is rapid: the reagent is added to a DNA solution, mixed and incubated for 5–10 minutes to allow complex formation, after which growth medium (which may contain serum and antibiotics) is added and the complexes are pipetted onto the cells. No post-transfection removal of complexes or medium change or addition is required, making the procedure highly suitable for high-throughput screening applications. The reagent is reported to consistently deliver outstanding transfection efficiencies with excellent reproducibility and minimal cytotoxicity, and is also available in bulk quantities for larger-scale work.

On arrival the reagent should be stored at 2–8°C. Cells should be 40–80% confluent at the time of transfection and incubated at 37°C with 5% CO2 to allow gene expression.