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Journal of Analytical Methods in Chemistry
Volume 2017 (2017), Article ID 4065892, 6 pages
https://doi.org/10.1155/2017/4065892
Research Article

Evaluation of Global Genomic DNA Methylation in Human Whole Blood by Capillary Electrophoresis UV Detection

1Department of Biomedical Sciences, University of Sassari, Sassari, Italy
2Department of Clinical and Experimental Medicine, University of Sassari, Sassari, Italy
3Department of Biomedical Sciences, College of Health Sciences, Qatar University, Doha 2713, Qatar
4Department of Clinical Pharmacology, College of Medicine and Public Health, Flinders University, Adelaide, SA, Australia

Correspondence should be addressed to Angelo Zinellu

Received 27 July 2017; Revised 16 October 2017; Accepted 30 October 2017; Published 3 December 2017

Academic Editor: Mohamed Abdel-Rehim

Copyright © 2017 Angelo Zinellu et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Alterations in global DNA methylation are implicated in various pathophysiological processes. The development of simple and quick, yet robust, methods to assess DNA methylation is required to facilitate its measurement and interpretation in clinical practice. We describe a highly sensitive and reproducible capillary electrophoresis method with UV detection for the separation and detection of cytosine and methylcytosine, after formic acid hydrolysis of DNA extracted from human whole blood. Hydrolysed samples were dried and resuspended with water and directly injected into the capillary without sample derivatization procedures. The use of a run buffer containing 50 mmol/L BIS-TRIS propane (BTP) phosphate buffer at pH 3.25 and 60 mmol/L sodium acetate buffer at pH 3.60 (4 : 1, v/v) allowed full analyte identification within 11 min. Precision tests indicated an elevated reproducibility with an interassay CV of 1.98% when starting from 2 μg of the extracted DNA. The method was successfully tested by measuring the DNA methylation degree both in healthy volunteers and in reference calf thymus DNA.