Abstract
Electroporation has been an established tool for DNA delivery into prokaryotic and eukaryotic cells, thus facilitating basic research studies and improving medical treatments. Here we describe its use for introduction of phage genomic DNA into Escherichia coli cells, including preparation of electrocompetent cells, electric pulse optimization and recovery of electrotransformed cells. The technique can also be adapted for other bacterial species.
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References
Kotnik T, Kramar P, Pucihar G, Miklavčič D, Tarek M (2012) Cell membrane electroporation—Part 1: the phenomenon. IEEE Electr Insul Mag 28:14–23
Satkauskas S, Ruzgys P, Venslauskas MS (2012) Towards the mechanisms for efficient gene transfer into cells and tissues by means of cell electroporation. Expert Opin Biol Ther 12:275–286
Lu Y-P, Zhang C, Lv FX, Bie XM, Lu Z-X (2012) Study on the electro-transformation conditions of improving transformation efficiency for Bacillus subtilis. Lett Appl Microbiol 55:9–14
Rivera AL, Magaña-Ortíz D, Gómez-Lim M, Fernández F, Loske AM (2014) Physical methods for genetic transformation of fungi and yeast. Phys Life Rev 11:184–203
Dower WJ, Chassy BM, Trevors JT, Blaschek HP (1992) Protocols for the transformation of bacteria by electroporation. In: Chang DC, Saunders JA, Chassy BM, Sowers AE (eds) Guide to electroporation and electrofusion. Academic Press, San Diego
Pucihar G, Krmelj J, Rebersek M, Batista Napotnik T, Miklavcic D (2011) Equivalent pulse parameters for electroporation. IEEE Trans Biomed Eng 58:3279–3288
Lelieveld HLM, Notermans S, de Haan SWH (eds) (2007) Food preservation by pulsed electric fields. Woodhead Publishing, Abington
Flisar K, Haberl Meglič S, Morelj J, Golob J, Miklavčič D (2014) Testing a prototype pulse generator for a continuous flow system and its use for E. coli inactivation and microalgae lipid extraction. Bioelectrochemistry 100:44–51
Kropinski AM, Mazzocco A, Waddell TE, Lingohr E, Johnson RP (2009) Enumeration of bacteriophages by double agar overlay plaque assay. In: Clokie MRJ, Kropinski AM (eds) Bacteriophages: volume 1: isolation, characterisation and interactions. Humana Press, New York
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Janež, N., Meglič, S.H., Flisar, K., Miklavčič, D., Peterka, M. (2019). Introduction of Phage Genome into Escherichia coli by Electroporation. In: Clokie, M., Kropinski, A., Lavigne, R. (eds) Bacteriophages. Methods in Molecular Biology, vol 1898. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8940-9_4
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DOI: https://doi.org/10.1007/978-1-4939-8940-9_4
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