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Electrotransformation of Saccharomyces cerevisiae.

Authors
  • Meilhoc, E1
  • Teissie, J2
  • 1 Laboratoire des Interactions Plantes-Microorganismes (LIPM), Université de Toulouse, INRA, CNRS, INSA, Castanet-Tolosan, France. , (France)
  • 2 Institut de Pharmacologie et de Biologie Structurale, Université de Toulouse, CNRS, UPS, Toulouse, France. [email protected] , (France)
Type
Published Article
Journal
Methods in molecular biology (Clifton, N.J.)
Publication Date
Jan 01, 2020
Volume
2050
Pages
187–193
Identifiers
DOI: 10.1007/978-1-4939-9740-4_21
PMID: 31468494
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Intact yeast cell transformation is easily achieved by gene electrotransfer (GET). The procedure is fast and efficient in terms of transformants/μg DNA. Yeast cells in exponential growth phase are washed, treated for a short period with dithiothreitol (DTT) and then mixed with the plasmid DNA in a buffer with a low conductivity. A single well defined electric pulsed is delivered. After a 1 h incubation in the growth medium without selection, transformants are obtained on a selective plate medium. After a short description of the present knowledge on the events affecting the yeast cell as a consequence of the pulsed electric field, a step-by-step protocol is reported for Saccharomyces cerevisiae.

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