The process of transfection involves introducing foreign DNA into cells. There are three different protocols for transfection: forward, reverse, and batch. The forward protocol involves splitting the cells a day before transfection and adding the jetPEI®/DNA complexes to the adherent or suspension cells. The reverse protocol is the best option for transfecting a gene pool, such as a DNA library.
In this protocol, the jetPEI®/DNA complexes are prepared or deposited in the wells before adding the cells. The complexes can remain stable for up to four hours. The batch protocol is used to prepare a homogeneous pool of transfected cells. The cells are transfected immediately after trypsinization while still in suspension to achieve this. This protocol is ideal for drug screening applications and allows for faster processing, one day faster than the forward protocol.
The jetPEI® and DNA complexes are formed using a water-soluble polymer that allows efficient transfection for up to four hours. This is in contrast to lipid-based reagents and calcium phosphate, which have a shorter time window for transfection. JetPEI® can be used to deliver genes to various adherent and non-adherent cell lines and primary cells. Over 550 publications using jetPEI® are available in the Polyplus-transfection Database, and specific transfection conditions for over 400 cell lines and primary cells can be found in their online database. JetPEI® was compared to several other popular transfection reagents and offered the best efficiency and low variability performance.