( .4 30 Endo-Free Anion-E...)Introducing,,,GenElute HP,,,Endotoxin-Free Plasmid,,,Maxiprep Kit

Transfection efficiencies into HuH-7 (human hepatoma) cells were compared among the different purification systems and reported in Figure 2. The results show that the sample purified using the GenElute HP Endotoxin-Free Plasmid Maxiprep Kit had the highest transfection efficiency when compared to the other commercially available kits.

Figure 2: Comparison of transfection efficiencies into HuH-7 cells using different purification systems with pCMV-SPORT-β-gal. The data show the average and standard deviations of six replicates from each sample prepared. HuH-7 cells were transfected using ESCORT II Transfection Reagent (Sigma Product Code L6037). The efficiency of the transfection was determined by measuring the b-galactosidase activity using the b-galactosidase Reporter Gene Activity Detection Kit (Sigma Product Code GALA-1KT).

Transfection of pCOP-Green-C into HuH-7 cells
Plasmid pCOP-Green-C is a mammalian expression vector that encodes the copepod green flourescence protein. HuH- 7 cells were seeded onto glass cover slips and then grown to 60-70% confluency. The cells were then transfected with 3 g of plasmid DNA using ESCORT II Transfection Reagent. Plasmid DNA was isolated from a GenElute HP Endotoxin- Free Maxiprep. At 72 hours post transfection, the glass cover slips were mounted onto microscope slides and the cells were observed under a fluorescence microscope. Panel A is a picture of the cells following transfection under bright field exposure. Panel B is the same area of cells under fluorescent exposure, which reveals a high transfection efficiency.

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