Quantitative Analysis of DNA Methylation by Bisulfite Sequencing

  • Vasily V. AshapkinEmail author
  • Lyudmila I. Kutueva
  • Boris F. Vanyushin
Part of the Methods in Molecular Biology book series (MIMB, volume 2138)


Changes in deoxyribonucleic acid (DNA) methylation are shown to occur with aging in mammals. Besides changes that seem to be essentially stochastic, methylation levels of certain CpG sites display a strong correlation with age. Collectively, methylation of such CpG sites could be used as “epigenetic clocks” to predict biological age. Numerous versions of the epigenetic clock have been proposed, all of them based on quantitative estimation of the methylation levels of individual CpG sites. Different methods were elaborated for quantitative measurements of DNA methylation, with the most reliable of these based on bisulfite treatment of DNA. We present here a protocol for assessment of the methylation levels of individual CpG sites in target DNA sequences by the direct sequencing of polymerase chain reaction (PCR) amplification products obtained from bisulfate-converted DNA.

Key words

Aging Epigenetics DNA methylation Bisulfite sequencing 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Vasily V. Ashapkin
    • 1
    Email author
  • Lyudmila I. Kutueva
    • 1
  • Boris F. Vanyushin
    • 1
  1. 1.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia

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