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Development of rapid microwave-mediated and low-temperature bacterial transformations

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Journal of Chemical Biology

Abstract

The introduction of exogenous DNA into Escherichia coli is a cornerstone of molecular biology. Herein, we investigate two new mechanisms for bacterial transformation involving either the use of microwave irradiation or a freeze–thaw protocol in liquid nitrogen. Ultimately, both methods afforded successful transfer of plasmid DNA into bacterial cells, with the freeze–thaw technique yielding efficiencies of ~105. More importantly, both techniques effectively eliminated the need for the preparation of competent cells.

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Acknowledgments

The authors would like to acknowledge the College of William & Mary for the funding, including a research award for D.D.Y. D.D.Y would like to thank the Jeffress Memorial Trust for the financial support. Additionally, J.C.M would like to thank the Howard Hughes Medical Institute for a research scholarship. Finally we would like to thank Dr. Mark Forsyth for providing the culture of M. smegmatis used in the experiments.

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Authors and Affiliations

  1. Department of Chemistry, The College of William & Mary, Williamsburg, VA, 23187, USA

    Valerie T. Tripp, Johnathan C. Maza & Douglas D. Young

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  1. Valerie T. Tripp
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  2. Johnathan C. Maza
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  3. Douglas D. Young
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Correspondence to Douglas D. Young.

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Tripp, V.T., Maza, J.C. & Young, D.D. Development of rapid microwave-mediated and low-temperature bacterial transformations. J Chem Biol 6, 135–140 (2013). https://doi.org/10.1007/s12154-013-0095-4

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  • DOI: https://doi.org/10.1007/s12154-013-0095-4

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