FIG. 1.

Example for a multivariate optimization layout. Schematic layout of different transfection variations (see Table 1) used for optimizing three transfection methods in one 96-well plate simultaneously. Variations of factor 1 are represented as wells filled with light (variation A) and dark (variation B) gray color. In the case of Lipofectamine LTX, the amount of DNA (μg) and PLUS reagent (μL) varied with X-tremeGene HP, the volume of transfection mix added to the wells varied; and with JetPrime the volume of reagent (μL) used varied. Variations of factor 2 are organized in well columns. At 24 and 48 h post transfection (hpt) incubation time points are organized in double rows. White wells held nontransfected cells used as controls for the cell viability assay. The crossed-out wells represent empty wells used for holding the Hoechst staining blank. The layout for the Matra-A transfection was similarly organized on an additional 96-well plate (layout not shown).