( nce and by quantitative PCR...)Commercial Plasmid Preparation Methods Yield DNA With Different Transfection Capabilities

nce and by quantitative PCR, we repeated transfections on VERO cells with increasing amounts of this DNA. Approximately 5 times more was needed to achieve a similar number of blue cells to that achieved with DNA pre p a red by method D, although b-galactosidase activity remained lower; still higher amounts of DNA resulted in a decrease in transfection efficiency (data not shown). Thus, plasmid prepared by method D not only gave the highest yield, it also gave optimal transfection efficiency. This was especially noteworthy in cells that are m o re difficult to transfect.

Preparation of DNA and subsequent transfections can be both time intensive and costly. Our study demonstrates that, while many time-saving methods for DNA purification are commercially available, DNA prepared by these methods varies in total yield and transfection efficiency. Plasmid DNA preparation using the Quantum Prep matrix produces high-quality DNA with optimal transfection efficiencies.

References
Felgner, P.L., et al., Lipofectin : A highly efficient, lipid-mediated DNA-transfection procedure, P roc. Natl. Acad. Sci. USA, 84, 74137417 (1987)

Geschwind, M. D., et al., Transfer of the nerve growth factor gene into cell lines and cultured neurons using a defective herpes simplex virus vector. Transfer of the NGF gene into cells by a HSV-1 vector, Brain Res. Mol. Brain Res., 24, 327335 (1994)

Heid, C. A., et al., Real time quantitative PCR, Genome Res., 6, 986994 (1996)

Rose, J. K., et al., A new cationic liposome reagent mediating nearly quantitative transfection of animal cells, BioTechniques, 10, 520525 (1991)

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