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Uses and Applications of X-tremeGENE Q2 Transfection Reagent

Despite the wide spectrum of cell types that can be transfected using currently available transfection reagents, there are significant differences in the efficiency of DNA delivery into different types of cell. In certain cell types, such as HeLa, K-562, and Jurkat, the broad-spectrum transfection reagents available today usually provide relatively low transfection efficiency and low-level protein expression. The X-tremeGENE Q2 Transfection Reagent (X-Q2 Reagent) from Roche Applied Science was specially designed and optimized for high-efficiency gene delivery and high-level protein expression in HeLa, K-562, and Jurkat cells.

X-tremeGENE Q2 Transfection Reagent provides exceptional transfection efficiency in K-562 cells; approximately 20 - 30% of cells are transfected (Figure 1).

Figure 1: K-562 cells transfected with the X-tremeGENE Q2 Transfection Reagent and the X-tremeGENE pM1-beta-Gal vector (Roche Applied Science) show strong beta-galactosidase staining. Cells were plated in a 24-well plate (1 X 105 /well) and transfected with a complex of plasmid DNA (2 g) and X-tremeGENE Q2 Transfection Reagent (4 l). 48 hours post transfection, cells were stained for beta-Gal expression using the beta-Gal Staining Set from Roche Applied Science, Cat. No. 1 828 673.

The level of beta-galactosidase (beta-Gal) expression within each K-562 cell can vary as seen here from intense blue to light blue. The level of beta-Gal expression obtained with the X-tremeGENE Q2 Transfection Reagent was several-fold higher than the next best competitor reagent recommended for the transfection of K-562 cells (data not shown), even when the same vector was used under optimized conditions for both reagents.

In Jurkat cells , the X-tremeGENE Q2 Transfection Reagent also produced beta-Gal expression that was several-fold higher than competitor reagents L-2 and DC (Figure 2). In these experiments, the X-tremeGENE pM1-beta-Gal vector was used with each transfection reagent (in other experiments, the ratios of DNA to transfection reagents and the total amount of complex added per cell were optimized for each reagent)


Figure 2: Jurkat cells transfected with the X-tremeGENE Q2 Transfection Reagent showed higher levels of beta-Gal expression than currently recommended competitor reagents L-2 or DC. Cells (4x105/ well) were transfected with the X-tremeGENE pM1-beta-Gal vector (Roche Applied Science) in a 24-well plate. In each case, the ratios of DNA / transfection reagent and the amount of complex used in each well following optimization were those recommended by the suppliers. The level of reporter gene expression was determined 48 hours post transfection, using the beta-Gal Reporter Gene Assay Kit (Roche Applied Science, Cat. No. 1 758 241).

Note: The level of expression of a transfected gene will vary with the specific cell type, number of cells transfected, quantity of plasmid DNA used, ratio of transfection reagent to DNA, type of expression vector, cell culture medium, and the culture conditions.



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