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Updated: Jun 24, 2026
Methodology for Accurate Detection of Mitochondrial DNA Methylation
Published on: May 20, 2018
Protocol for mitochondrial DNA extraction, purity evaluation, and global methylation assessment via LC-MS/MS
Argyri Chroni1, Theodora Alexiou2, John Zafeiropoulos3
1Biology Laboratory, School of Science and Technology, Hellenic Open University, Patras 26335, Greece; School of Health Sciences, Faculty of Medicine, Department of Internal Medicine, University of Patras, and "Olympion" General Clinic, Department of Hematology, Patras, Greece.
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Established epigenetic mechanisms, including nuclear (n)DNA methylation, are also known to regulate the mitochondrial (mt) genome despite its lack of histones. However, studies referring to evaluations of mtDNA methylation are often influenced by technical and biological confounders. We present an optimized protocol for extracting mtDNA from peripheral blood and bone marrow mononuclear cells, followed by steps for qualitative assessment to exclude nDNA contaminations and a downstream quantitative analysis of the mtDNA global methylation profile using liquid chromatography combined with mass spectrometry.
