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Linking Microbes to Their Genes at Single Cell Level with Direct-geneFISH

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Fluorescence In-Situ Hybridization (FISH) for Microbial Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2246))

Abstract

Direct-geneFISH is a Fluorescence In Situ Hybridization (FISH) method that directly links gene presence, and thus potential metabolic capabilities, to cell identity. The method uses rRNA-targeting oligonucleotide probes to identify cells and dsDNA polynucleotide probes carrying multiple molecules of the same fluorochrome to detect genes. In addition, direct-geneFISH allows quantification of the cell fraction carrying the targeted gene and the number of target genes per cell. It can be applied to laboratory cultures, for example, enrichments and phage infections, and to environmental samples. This book chapter describes the main steps of the direct-geneFISH protocol: probe design and synthesis, the “core” direct-geneFISH protocol and lastly, microscopy and data analysis.

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References

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Author information

Authors and Affiliations

  1. Department of Environmental Technology, Technische Universität Berlin, Berlin, Germany

    Jimena Barrero-Canosa

  2. Institute for Chemistry and Biology of the Marine Environment, Oldenburg, Germany

    Cristina Moraru

Authors
  1. Jimena Barrero-Canosa
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  2. Cristina Moraru
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Corresponding author

Correspondence to Cristina Moraru .

Editor information

Editors and Affiliations

  1. LEPABE – Laboratory for Process Engineering Environment, Biotechnology and Energy, Department of Chemical Engineering, University of Porto, Porto, Portugal

    Nuno F. Azevedo

  2. INIAV – National Institute for Agrarian and Veterinarian Research, Rua dos Lagidos, Lugar da Madalena, Vairão, CEB - Centre of Biological Engineering, University of Minho, Campus de Gualtar, Braga, LEPABE – Laboratory for Process Engineering Environment, Biotechnology and Energy, Department of Chemical Engineering, University of Porto, Vila do Conde, Portugal

    Carina Almeida

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Barrero-Canosa, J., Moraru, C. (2021). Linking Microbes to Their Genes at Single Cell Level with Direct-geneFISH. In: Azevedo, N.F., Almeida, C. (eds) Fluorescence In-Situ Hybridization (FISH) for Microbial Cells. Methods in Molecular Biology, vol 2246. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1115-9_12

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  • DOI: https://doi.org/10.1007/978-1-0716-1115-9_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1114-2

  • Online ISBN: 978-1-0716-1115-9

  • eBook Packages: Springer Protocols

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