Versatile Transfection Reagent Offers Low Toxicity and Consistent,,,Performance ( IH 3T3 cell lines ( Figure 2...)

Versatile Transfection Reagent Offers Low Toxicity and Consistent,,,Performance

IH 3T3 cell lines (Figure 2).⁵ This study used the b-galactosidase control plasmid and varying amounts of lipid reagents for transfections.

Figure 2

The b-galactosidase activity of transfected cell lysates indicated transfection efficiency. To determine toxicity, for each volume of transfection reagent used in each cell line, we measured the concentration of cellular proteins (data not shown). LipoTAXI reagent gave the highest transfection efficiency and lowest toxicity in all three cell lines.

Transfection Efficiencies: Effect of Mycoplasma Contamination

When standardization studies are performed to determine optimized amounts of LipoTAXI transfection reagent and DNA, low efficiencies or atypical transfection profiles may be observed. This may be due to mycoplasma contaminating the cell lines, since antibiotic-resistant mycoplasma can exist in cell cultures without noticeable visual effects on cells.

Figure 3

In one laboratory, optimization of LipoTAXI reagent using the b-galactosidase reporter plasmid in HeLa cells showed uncharacteristically low efficiencies.⁶ Stratagenes Mycoplasma PCR Primer Set,*^, which provides a quick, sensitive assay for detecting mycoplasma in cell cultures, was used to test the HeLa cells for contamination. Cell-free supernatant samples were prepared, then analyzed, according to the protocol provided with the primer set. Results indicated that mycoplasma contamination was present in the HeLa cells at the time atypical optimization profiles were observed (Figure 3, lane 2). After the contaminated cells were destroyed and the laboratory
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