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Preparation of Competent Cells for High-Efficiency Plasmid Transformation of Escherichia coli

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cDNA Library Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 69))

Abstract

Transformation of Escherichia coli was first described by Mandel and Higa (1), who reported that E. coli cells, after treatment with calcium chloride, can take up bacteriophage λ DNA and produce viable phage particles. The conditions for the transfer of exogenous DNA into E. coli have been examined in detail in studies of bacteriophage transfection, genetic transformation, and plasmid transformation. Modifications that improve the efficiency of transformation include prolonged exposure of cells to CaCl2 (2), substitution of Ca2+ ions by other cations, such as Rb+ (3), Mn2+, and K+, and addition of other compounds, such as dimethyl sulfoxide (DMSO), dithiothreitol, and cobalt hexaminechloride (4).

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References

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© 1997 Humana Press Inc.

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Nakata, Y., Tang, X., Yokoyama, K.K. (1997). Preparation of Competent Cells for High-Efficiency Plasmid Transformation of Escherichia coli. In: Cowell, I.G., Austin, C.A. (eds) cDNA Library Protocols. Methods in Molecular Biology™, vol 69. Humana Press. https://doi.org/10.1385/0-89603-383-X:129

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  • DOI: https://doi.org/10.1385/0-89603-383-X:129

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-383-2

  • Online ISBN: 978-1-59259-555-6

  • eBook Packages: Springer Protocols

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