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High-Efficiency Transfections Achieved with New Low-Toxicity Reagent

ance in protocols that show actual experimental conditions. In most of these comparative tests, GeneJammer reagent was more effective than the other transfection products; it showed high efficiencies in conditions that are typically problematic, such as in the presence of serum or in transfections of primary cells. Conveniently, GeneJammer reagent does not require reconstitution before use so handling time is minimized.

Optimizing Conditions

Before studies were compared, we optimized transfection conditions for GeneJammer and each of the five most common transfection reagents (products a, b, c, d, and e). We used the pCMV b-gal reporter plasmid (0.4 g/well) to transfect CHO-K1, COS-7, 293, HeLa, human umbilical arterial endothelial cells (HUAEC), and human umbilical vein endothelial cells (HUVEC) in 24-well plates. For optimization, we used four different concentrations of GeneJammer reagent (3, 6, 9, and 12 l per 1 g of DNA) and three concentrations of the competitive reagents within the ranges recommended by each manufacturer. Then the experiments were carried out according the manufacturers protocols.

Transfection assays with GeneJammer reagent included serum-free and serum-containing medium; the competitive reagents were used in the absence of serum. The in situ transfection efficiency was evaluated by the b-galactosidase histochemical staining assay; cell viability was calculated by the trypan blue exclusion assay. For each cell line, optimization was defined as the volume of each reagent that provided the highest transfection efficiency and the lowest toxicity. These optimized values were confirmed in three independent trials per cell line.

Comparing Transfection Efficiencies

Figure 1 shows the transfection efficiency of GeneJammer reagent in the presence and absence of serum, compared with the competitive rea
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