A simple, highly selective, and sensitive method has been developed to quantify methylation of DNA extracted from human peripheral blood mononuclear cells. Assay has been performed at nucleobases level. Cytosine and 5-methylcytosine DNA content has been detected by gas chromatography-mass spectrometry using [2-13 C]cytosine and [2-13 C]5-methylcytosine as internal standards. The methylation level has been calculated as 5-methylcytosine/total cytosine ratio. The working range selected on calibration curve, obtained by evaluation of standards and matrix-added standards measurements, is suitable for 5 μg DNA analysis. In this range, healthy human DNA methylation percentage is within 5–6%.
|Titolo:||Measurement of DNA methylation using stable isotope dilution and gas chromatography-mass spectrometry.|
|Parole Chiave:||Cytosine; 5-Methylcytosine; Stable isotopes; DNA methylation; Gas chromatography-mass spectrometry.|
|Settore Scientifico Disciplinare:||Settore BIO/09 - Fisiologia|
|Data di pubblicazione:||gen-2005|
|Digital Object Identifier (DOI):||10.1016/j.ab.2004.09.034|
|Appare nelle tipologie:||01 - Articolo su periodico|