Abstract
Electroporation is now being used to transfer into cells a variety of macromolecules, including DNA, RNA, protein, fluorescent dyes, and some chemotherapeutic agents. With electrotransformation, as many as 80% of the cells receive exogenous DNA (1). Chemical and natural methods of DNA transfer into bacteria are also effective for transforming bacteria, but they are more cumbersome and time-consuming, and generally less efficient than electrotransformation.
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References
Dower, W. J., Miller, J. F., and Ragsdale, C. W. (1988) High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 16, 6127–6145.
Zell, R. and Fritz, H.-J. (1987) DNA mismatch-repair in Escherichia coli counteracting the hydrolytic deamination of 5-methyl-cytosine residues. EMBO J. 6, 1809–1815.
Deng, W. P. and Nickoloff, J. A. (1992) Site-directed mutagenesis of virtually any plasmid by eliminating a unique site. Anal. Biochem. 200, 81–88.
Ray, F. A. and Nickoloff, J. A. (1992) Site-specific mutagenesis of almost any plasmid using a PCR-based version of unique-site elimination. Biotechniques 13, 342–346.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Antonov, P. A., Maximova, V. A., and Pancheva, R. P. (1993) Heat shock and osmotically dependent steps by DNA uptake in Escherichia coli after electroporation. Biochim. Biophys. Acta 1216, 286–288.
Shigekawa, K. and Dower, W. J. (1988) Electroporation of eukaryotes and prokaryotes: a general approach to the introduction of macromolecules into cells. Biotechniques 6, 742–751.
Ray, F. A., Miller, E. M., and Nickoloff, J. A. (1994) Adapting unique site elimination site-directed mutagenesis for marker rescue and domain replacement. Anal. Biochem. (in press).
Nickoloff, J. A. (1994) Sepharose spin column chromatography: a fast, nontoxic replacement for phenol:chloroform extraction/ethanol precipitation. Mol. Biotechnol. 1, 105–108.
Siguret, V., Ribba, A.-S., Cherel, G., Meyer, D., and Pietu, G. (1994) Effect of plasmid size on transformation efficiency by electroporation of Escherichia coli DH5α. Biotechniques 16, 422–424.
Conley, E. C. and Saunders, J. R. (1984) Recombination-dependent recircularization of linearized pBR322 plasmid DNA following transformation of Escherichia coli. Mol. Gen. Genet. 194, 211–218.
Taghian, D. G. and Nickoloff, J. A. Subcloning strategies and protocols, in Basic DNA and RNA Protocols (Harwood, A., ed.), Humana, Totowa, NJ (in press).
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© 1995 Humana Press Inc.
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Miller, E.M., Nickoloff, J.A. (1995). Escherichia coli Electrotransformation. In: Nickoloff, J.A. (eds) Electroporation Protocols for Microorganisms. Methods in Molecular Biology™, vol 47. Humana Press. https://doi.org/10.1385/0-89603-310-4:105
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DOI: https://doi.org/10.1385/0-89603-310-4:105
Publisher Name: Humana Press
Print ISBN: 978-0-89603-310-8
Online ISBN: 978-1-59259-534-1
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