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Colony PCR

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1620))

Abstract

Escherichia coli and Saccharomyces cerevisiae are currently the two most important organisms in synthetic biology. E.coli is almost always used for fundamental DNA manipulation while yeast is the simplest host system for studying eukaryotic gene expression and performing large scale DNA assembly. Yeast expression studies may also require altering of the chromosomal DNA by homologous recombination. All these studies require the verification of the expected DNA sequence and the fastest method of screening is colony PCR, which is direct PCR of DNA in cells without prior DNA purification. Colony PCR is hampered by the difficulty of releasing DNA into the PCR mix and the presence of PCR inhibitors. We hereby present one protocol for E. coli and two protocols for S. cerevisiae differing in efficiency and complexity as well as an overview of past and possible future developments of efficient S. cerevisiae colony PCR protocols.

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Acknowledgment

This work was supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI).

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Authors and Affiliations

  1. Centre of Molecular and Environmental Biology (CBMA), Department of Biology, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal

    Flávio Azevedo, Humberto Pereira & Björn Johansson

Authors
  1. Flávio Azevedo
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  2. Humberto Pereira
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  3. Björn Johansson
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Corresponding author

Correspondence to Björn Johansson .

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Editors and Affiliations

  1. CEB—Centre of Biological Engineering, University of Minho, Braga, Portugal

    Lucília Domingues

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Azevedo, F., Pereira, H., Johansson, B. (2017). Colony PCR. In: Domingues, L. (eds) PCR. Methods in Molecular Biology, vol 1620. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-7060-5_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7060-5_8

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4939-7059-9

  • Online ISBN: 978-1-4939-7060-5

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