Critical factors for successful RNAi experiments in primary cells and hard to transfect cell lines

The amaxa Nucleofector® Technology is a well established method for effective, non-viral transfection of any nucleic acid substrate into hard-to-transfect cells, especially suspension and primary cells. With the Nucleofector® 96-well Shuttle® system high throughput applications such as siRNA-library screenings have become amenable for the first time in these cell types. This renders target validation and identification possible in cell types highly relevant for biomedical research. Here we discuss critical factors for setting up RNAi experiments using Nucleofection®. We focused on relevant cells poorly accessible with reagent-based transfection methods and present data showing the efficient RNAi-mediated gene knockdown in Jurkat cells T-cells.