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A Capillary Electrophoresis UV Detection-Based Method for Global Genomic DNA Methylation Assessment in Human Whole Blood

  • Angelo ZinelluEmail author
  • Elisabetta Sotgiu
  • Salvatore Sotgia
  • Ciriaco Carru
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1972)

Abstract

Quantitative analysis of DNA methylation patterns is of special importance in several developmental and pathological situations. The development of simple and robust methods to assess DNA methylation is required to facilitate its measurement and interpretation in clinical practice. We describe a highly reproducible CE-UV method for the separation and detection of cytosine and methylcytosine, after formic acid hydrolysis of DNA extracted from human whole blood. Hydrolyzed samples were dried and successively dissolved with water and then 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 a baseline analytes separation within 12 min. Precision tests indicated an elevated reproducibility with an inter-assay CV of 1.98%.

Key words

Capillary electrophoresis DNA methylation 5-Methylcytosine 

Notes

Acknowledgements

This work was supported by CNR—DSBM UNISS Flagship InterOmics (cod. PB05).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Angelo Zinellu
    • 1
    Email author
  • Elisabetta Sotgiu
    • 1
  • Salvatore Sotgia
    • 1
  • Ciriaco Carru
    • 1
  1. 1.Department of Biomedical SciencesUniversity of SassariSassariItaly

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