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Application of Nucleic Acid Mimics in Fluorescence In Situ Hybridization

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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

Traditionally, RNA and DNA probes are used in fluorescence in situ hybridization (FISH) methods for microbial detection and characterization of communities’ structure and diversity. However, the recent introduction of nucleic acid mimics (NAMs) has improved the robustness of the FISH methods in terms of sensitivity and specificity. Several NAMs have been used, of which the most relevant are peptide nucleic acid (PNA), locked nucleic acids (LNA), 2′-O-methyl RNA (2′OMe), and phosphorothioates (PS). In this chapter, we describe a protocol using PNA and LNA/2’OMe probes for microbial detection by FISH, pointing out the differences between them. These protocols are easily adapted to different microorganisms and different probe sequences.

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Acknowledgements

This work was financially supported by (a) Base Funding—UIDB/00511/2020 of the Laboratory for Process Engineering, Environment, Biotechnology and Energy—LEPABE—funded by national funds through the FCT/MCTES (PIDDAC); (b) Projects POCI-01-0145-FEDER-016678 (Coded-FISH), POCI-01-0145-FEDER-03043 (CLASInVivo) and POCI-01-0145-FEDER- 028659 (NAM4toxins), funded by FEDER funds through COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI), and by national funds (PIDDAC) through FCT/MCTES.

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Authors and Affiliations

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

    Ricardo Oliveira & Andreia S. Azevedo

  2. INIAV – National Institute for Agrarian and Veterinarian Research, Rua dos Lagidos, Lugar da Madalena, Vairão, Vila do Conde, Portugal

    Ricardo Oliveira

  3. i3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal

    Andreia S. Azevedo

  4. IPATIMUP – Institute of Molecular Pathology and Immunology of the University of Porto, University of Porto, Porto, Portugal

    Andreia S. Azevedo

  5. CEB – Centre of Biological Engineering, University of Minho, Braga, Portugal

    Andreia S. Azevedo

  6. FMDUP – Faculty of Dental Medicine, University of Porto, Porto, Portugal

    Luzia Mendes

Authors
  1. Ricardo Oliveira
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  2. Andreia S. Azevedo
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  3. Luzia Mendes
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Corresponding author

Correspondence to Luzia Mendes .

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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Oliveira, R., Azevedo, A.S., Mendes, L. (2021). Application of Nucleic Acid Mimics in Fluorescence In Situ Hybridization. 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_5

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  • DOI: https://doi.org/10.1007/978-1-0716-1115-9_5

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  • Publisher Name: Humana, New York, NY

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  • Online ISBN: 978-1-0716-1115-9

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