FAQ: Generalities about transfection

In contrast to many other transfection reagents, all our products are compatible with serum therefore complexes can be directly added to cells in serum-containing medium.

None of our products transfection efficiency is affected by the presence of antibiotics. For example, the routine validation protocol for each new batch of jetPRIME® is always performed in serum and antibiotics containing medium.

Many cell culture media are used to grow and maintain mammalian cells and some of them may inhibit cationic polymer-mediated transfection. For more information about cell medium compatibility with transfection, contact our delivery experts at support@polyplus-transfection.com.

Passage numbers will affect transfection performance. If cells have been in culture for a long time (over 20 passages), transfection efficiency will decrease and we recommend starting a new culture with a fresh tube of cells from liquid nitrogen. On the other hand, cells should be passaged at least twice after thawing before starting transfection experiments.

Cell density will affect transfection performance. Generally, we recommend transfecting cells at 60% to 80% confluency. Please refer to the transfection reagent protocol for the recommended number of cells to seed according to the culture plate format.

The ratio corresponds the number of microliters of transfection reagent to use per microgram of nucleic acids. For example, a 1:2 DNA to jetPRIME® ratio means 2 µl jetPRIME® per µg DNA.

The first step of optimization is to increase the DNA amount up to 1.5 fold. We also recommend increasing the nucleic acid: transfection reagent ratio. Another way to increase transfection efficiency consists in implementing a gentle centrifugation of the culture plate (5 min at 210 g) after complexes addition.