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Methylation-Specific PCR

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DNA Methylation Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1708))

Abstract

Cytosine methylation is a DNA modification generally associated with transcriptional silencing. Factors that regulate methylation have been linked to human disease, yet how they contribute to malignancies remains largely unknown. Methylation of DNA can change the functional state of regulatory regions, but does not change the Watson–Crick base pairing of cytosine. Moreover, sequence symmetry of CpGs enables propagation of the methyl mark through cell division. This potential for inheritance coupled with the fact that DNA methylation patterns change during development and disease partially explains the interest in DNA methylation as a memory module. DNA methylation analysis also provides an opportunity to discover new and more powerful biomarkers that can help in clinical practice.

Methylation-Specific PCR (MSP) is likely the most widely used technique to study DNA methylation of a locus of interest. MSP can rapidly detect the methylation status of any group of CpG sites within a CpG island, not requiring methylation-sensitive restriction enzymes. It also requires minute amounts of DNA, is very sensitive as it can detect <0.1% of methylated alleles in a specific locus, and can be used in different samples, including bodily fluids, and paraffin-embedded samples.

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Acknowledgments

This work was supported by Federal funds through Programa Operacional Temático Factores de Competitividade (COMPETE) with co-participation from the European Community Fund (FEDER) and by national funds through Fundação para a Ciência e Tecnologia (FCT) under the project EXPL/BIM-ONC/0556/2012 (CJ). João Ramalho-Carvalho is supported by a fellowship from FCT (SFRH/BD/71293/2010).

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Authors and Affiliations

  1. Cancer Biology and Epigenetics Group; Research Center (CI-IPOP), Portuguese Oncology Institute of Porto (IPO Porto), Porto, Portugal

    João Ramalho-Carvalho, Rui Henrique & Carmen Jerónimo

  2. Biomedical Sciences Graduate Program, Institute of Biomedical Sciences Abel Salazar, University of Porto (ICBAS-UP), Porto, Portugal

    João Ramalho-Carvalho

  3. Department of Pathology, Portuguese Oncology Institute of Porto (IPO Porto), Porto, Portugal

    Rui Henrique

  4. Department of Pathology and Molecular Immunology, Institute of Biomedical Sciences Abel Salazar–University of Porto (ICBAS-UP), Porto, Portugal

    Rui Henrique & Carmen Jerónimo

  5. Portuguese Oncology Institute of Porto, Research Center-LAB 3, F Bdg, 1st floor, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal

    Carmen Jerónimo

Authors
  1. João Ramalho-Carvalho
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  2. Rui Henrique
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  3. Carmen Jerónimo
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Corresponding author

Correspondence to Carmen Jerónimo .

Editor information

Editors and Affiliations

  1. Laboratory for Epigenetics and Environment, Centre National de Recherche en Génomique Humaine, CEA—Institut de Biologie Francois Jacob, Evry, France

    Jörg Tost

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Ramalho-Carvalho, J., Henrique, R., Jerónimo, C. (2018). Methylation-Specific PCR. In: Tost, J. (eds) DNA Methylation Protocols. Methods in Molecular Biology, vol 1708. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7481-8_23

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  • DOI: https://doi.org/10.1007/978-1-4939-7481-8_23

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7479-5

  • Online ISBN: 978-1-4939-7481-8

  • eBook Packages: Springer Protocols

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