Animal Cell Electroporation and Electrofusion Protocols by Jac A. Nickoloff

By Jac A. Nickoloff

Harvard collage, Boston, Massachusetts. handbook of protocols within the electroporation of proteins and DNA into fish, insect, and mammalian cells. For researchers. Plastic comb spiral binding. sixty five individuals, 28 U.S.

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Extra info for Animal Cell Electroporation and Electrofusion Protocols (Methods in Molecular Biology)

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Square- Wave Generators Square-wave pulses appearto have advantagesfor certain applications, such as transfection of mammalian cell lines and plant protoplasts, though no generalization can be made. Each cell line needs to be individually investigated to determine whether use of square-wave pulses would be advantageous. In general, square waves do not appear to result in higher transformation yields for bacteria, although there are some protocols that give good results (6; Xing Xin, Texas Heart Institute, personal communication).

Effect of electric parameters ET on pRSVgpt transfection of HeLa cells. The number of successfully transfected HeLa cells with pRSVgpt plasmid by electroporatton 1s plotted as a function of the product of peak field strength E and decay half-time %112 of an applied exponential pulse. ). found to be a linear function of either E or T, when the other parameter was held constant. 6 kV/cm. Interestingly, the transfection efficiency decreases with increasing delay between repeating pulses, indicating that DNA is collected on the cell surface and driven through the electropores by electrophoretic force, in agreement with previous experiments using two consecutive pulses to increase transfection yield (8).

1988) Introductton and expression of DNA molecules in eukaryotic cells by electroporation (published erratum appears in BioTechmques 6,853) BioTechnrques 6,650-660. 3. , and Teissie, J. (1992) Fast kinetics studies of Escherichia coli electrotransformation. Eur. J Biochem. 209,431-436. 4. Morgan, W. F. and Wmegar, R. A. (1990) The use of restriction endonucleases to study the mechamsms of chromosome damage, in Chromosomal Aberrations’ Basic and Applied Aspects (Obe, G. and Natarajan, A. ), Springer-Verlag, Berlin, pp.

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