Quantification of global DNA methylation by capillary electrophoresis and mass spectrometry.

Full Abstract

Two approaches for the evaluation of the relative degree of global DNA methylation through the quantification of 2' deoxynucleosides are described. Detection and quantification of 5-methyl 2'-deoxycytidine in genomic DNA is performed using both high-performance capillary electrophoresis (HPCE) with UV-Vis detection or liquid chromatography with electrospray ionization mass spectrometric detection (LC-ESI/MS). Treatment of genomic DNA with a ribonuclease and generation of nucleosides through enzymatic hydrolysis notably increases the specificity of both techniques. Both approaches have been demonstrated to be highly specific and sensitive, being useful for the rapid quantification of the degree of global DNA methylation and its exploitation for the analysis of poorly purified and/or concentrated DNA samples, such as tumor biopsies.

Author information

Author/s: Berdasco, María (M); Fraga, Mario F (MF); Esteller, Manel (M);

Affiliation: Epigenetics Group, Molecular Pathology Program, Spanish National Cancer Center (C.N.I.O.), Madrid, Spain.

Journal and publication information

Publication Type: Journal Article

Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol), published in United States. (Language: eng)

Reference: 2009-; vol 507 (issue ) : pp 23-34

Dates: Created 2008/11/06; Completed 2009/03/03;

PMID: 18987803, status: MEDLINE (last retrieved date: 3/10/2009)

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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Associated Chemicals: DNA, Neoplasm (0) ; DNA (9007-49-2)

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