HiPerFect Transfection Reagent (1 ml)

QIAGEN supplies a transfection reagent designed for delivery of siRNA or miRNA into eukaryotic cells — adherent and suspension cultures — in 24-well plate formats. HiPerFect Transfection Reagent should be stored at 2–8 °C upon arrival. A product handbook, safety data sheets, and technical-assistance contact information are available at www.qiagen.com.

The accompanying protocol outlines cell preparation, complex formation, and incubation under standard growth conditions, typically 37 °C and 5% CO2. For adherent cells, 0.4–1.6 × 10^5 cells are seeded per well in 0.5 ml medium, with siRNA diluted to a final 5 nM and 3 µl of reagent per well. For suspension lines such as Jurkat and K562, 2 × 10^5 cells per well are plated in 100 µl medium and combined with 750 ng siRNA (100 nM final) and 6 µl of reagent. Differentiated macrophage lines such as THP-1 are seeded at 2 × 10^4 cells per well 24 h before transfection in 0.5 ml medium containing a differentiation-inducing agent at 100 ng/ml PMA.

Transfection complexes form during 5–10 min of incubation at room temperature (15–25 °C) and are added drop-wise onto the cells. Gene silencing is monitored 6–72 h after transfection, depending on the experimental setup. Suggested siRNA-to-reagent ratios for optimisation range from 7.5 ng to 750 ng of siRNA with 1.5 µl to 9 µl of reagent. A regularly updated list of successfully transfected cell lines is available through the Transfection Cell Database at www.qiagen.com/TransfectionTools.