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Electron Microscope Detection of 5-Methylcytosine on DNA and RNA

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Epitranscriptomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1870))

Abstract

5-Methylcytosine is the major epigenetic modification occurring on DNA. It is known to be involved not only in gene expression regulation but also in the control of chromatin structure. However, this modification is also found on different types of RNA, including mRNA. Generally, biomolecular techniques are applied for studying the epigenetic profile of nucleic acids. Here, we describe the ultrastructural detection of 5-methylcytosine as an unusual approach to localize this modification on chromatin regions and/or RNA single molecules. This tool requires a careful sample preparation to preserve antigen epitopes that will be revealed immunocytochemically by a specific anti-5-methylcytosine antibody. The multiple staining procedures that can be adopted allow the identification of both DNA or RNA. A semiquantitative analysis can also be carried out.

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Acknowledgments

The authors would like to thank Ms. Francine Flach for excellent technical skill in preparing the ultrathin sections.

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Authors and Affiliations

  1. Laboratory of Cell Biology and Neurobiology, Department of Biology and Biotechnology, University of Pavia, Pavia, Italy

    Irene Masiello & Marco Biggiogera

Authors
  1. Irene Masiello
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  2. Marco Biggiogera
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Corresponding author

Correspondence to Marco Biggiogera .

Editor information

Editors and Affiliations

  1. Department of Pathology, Yale University School of Medicine, New Haven, CT, USA

    Narendra Wajapeyee

  2. Yale Cancer Center, Yale University School of Medicine, New Haven, CT, USA

    Romi Gupta

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Masiello, I., Biggiogera, M. (2019). Electron Microscope Detection of 5-Methylcytosine on DNA and RNA. In: Wajapeyee, N., Gupta, R. (eds) Epitranscriptomics. Methods in Molecular Biology, vol 1870. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8808-2_12

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  • DOI: https://doi.org/10.1007/978-1-4939-8808-2_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8807-5

  • Online ISBN: 978-1-4939-8808-2

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