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Bacteria and Yeast Colony PCR

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2967))

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Abstract

The bacteria Escherichia coli and the yeast 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 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 by 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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Acknowledgments

This work was supported by the Fundação para a Ciência e Tecnologia Portugal (FCT) through Project FatVal PTDC/EAM-AMB/032506/2017 funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020 – Programa Operacional Competitividade e Internacionalizacão (POCI). CBMA was supported by the strategic program UIDB/04050/2020 funded by national funds through the FCT I.P. Humberto Pereira acknowledges FCT for the Ph.D. scholarship, SFRH/BD/148722/2019.

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Authors and Affiliations

  1. CBMA - Centre of Molecular and Environmental Biology, Department of Biology, University of Minho, Braga, Portugal

    Humberto Pereira, Paulo César Silva & Björn Johansson

Authors
  1. Humberto Pereira
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  2. Paulo César Silva
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  3. Björn Johansson
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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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Pereira, H., Silva, P.C., Johansson, B. (2023). Bacteria and Yeast Colony PCR. In: Domingues, L. (eds) PCR. Methods in Molecular Biology, vol 2967. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3358-8_17

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  • DOI: https://doi.org/10.1007/978-1-0716-3358-8_17

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3357-1

  • Online ISBN: 978-1-0716-3358-8

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