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Nucleotide-Level Profiling of m5C RNA Methylation

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Post-Transcriptional Gene Regulation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1358))

Abstract

Mapping the position and quantifying the level of 5-methylcytosine (m5C) as a modification in different types of cellular RNA is an important objective in the emerging field of epitranscriptomics. Bisulfite conversion has long been the gold standard for detection of m5C in DNA but it can also be applied to RNA. Here, we detail methods for bisulfite treatment of RNA, locus-specific PCR amplification and detection of candidate sites by sequencing on the Illumina MiSeq platform.

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Acknowledgements

We thank Wenjia Qu for helpful suggestions for the MiSeq library preparation protocol. We also thank Ulrike Schumann for helpful suggestions on this manuscript. This work was supported by an NHMRC grant (APP1061551) and a Senior Research Fellowship (514904) awarded to TP.

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Authors and Affiliations

  1. Genome Biology Department, The John Curtin School of Medical Research (JCSMR), The Australian National University, Acton, Canberra, ACT, Australia

    Tennille Sibbritt, Andrew Shafik & Thomas Preiss

  2. Genomics and Epigenetics Department, Garvan Institute of Medical Research, Darlinghurst, Sydney, NSW, Australia

    Susan J. Clark

  3. Victor Chang Cardiac Research Institute, Darlinghurst, Sydney, NSW, Australia

    Thomas Preiss

Authors
  1. Tennille Sibbritt
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  2. Andrew Shafik
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  3. Susan J. Clark
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  4. Thomas Preiss
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Corresponding author

Correspondence to Thomas Preiss .

Editor information

Editors and Affiliations

  1. Laboratory of Translational Genomics, Centre for Integrative Biology, University of Trento, Mattarello, Trento, Italy

    Erik Dassi

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Sibbritt, T., Shafik, A., Clark, S.J., Preiss, T. (2016). Nucleotide-Level Profiling of m5C RNA Methylation. In: Dassi, E. (eds) Post-Transcriptional Gene Regulation. Methods in Molecular Biology, vol 1358. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3067-8_16

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  • DOI: https://doi.org/10.1007/978-1-4939-3067-8_16

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3066-1

  • Online ISBN: 978-1-4939-3067-8

  • eBook Packages: Springer Protocols

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