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Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection,,,of Mammalian Cells


Mycoplasma PCR Primer Set for fast and easy detection of mycoplasma contamination

Leslie S. Arrington
Yale University

D. R. Staggs
Genetic Applications LLC

Heinz Miller
Stratagene

Mycoplasma contamination of mammalian cell cultures can substantially reduce the potency of transfection reagents.1 Cells must be free of mycoplasma contamination before the optimal amounts of LipoTAXI transfection reagent and DNA can be accurately determined. The use of Stratagenes Mycoplasma PCR Primer Set*, provides assurance that cells are free of mycoplasma contamination and able to maximally express transfected DNA.

The efficient introduction of foreign DNA into mammalian cells is vital to many areas of biological research. The ability to study the effect of introducing a gene or oligonucleotide of interest in a cell line may depend on the effectiveness of the transfection reagent. Stratagenes LipoTAXI transfection reagent can be used to achieve efficient transfections in a wide variety of mammalian cells. Using simple protocols, researchers can determine the optimal combination of LipoTAXI transfection reagent and plasmid DNA for a particular cell type.2 However, mycoplasma contamination can interfere with transfection using lipid, DEAE-dextran, CaPO4 and adenovirus-mediated methods.1 The resulting low transfection efficiencies may go undetected since antibiotic-resistant mycoplasma can exist in cell cultures without noticeable visual effects on the cells.3

In the typical protocol for optimizing transfection conditions, the amount of LipoTAXI reagent is increased at various concentrations of plasmid DNA that encodes for a reporter gene. Expression of the reporter
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