High-performance capillary electrophoretic method for the quantification of 5-methyl 2'-deoxycytidine in genomic DNA: application to plant, animal and human cancer tissues.
A new approach to the evaluation of the relative degree of genomic DNA methylation through the quantification of 2'-deoxynucleosides is proposed. Detection and quantification of 5-methyl 2'-deoxycytidine in genomic DNA has been performed using micellar high-performance capillary electrophoresis (HPCE) with UV-Vis detection. This approach has been demonstrated to be more sensitive and specific than other HPCE methods for the quantification of DNA methylation degree and also to be faster than other HPLC-based methods. The detection and quantification of nucleosides through enzymatic hydrolyses notably increases the specificity of the technique and allows its exploitation in the analysis of poorly purified and/or concentrated DNA samples such as those obtained from meristematic plant regions and paraffin-embedded tissues.
Fraga
Mario F
MF
Laboratorio de Fisiología Vegetal, Dpt. B.O.S., Facultad de Biología, Universidad de Oviedo, Oviedo, Spain. mffraga@CNIO.es
Uriol
Esther
E
Borja Diego
L
L
Berdasco
María
M
Esteller
Manel
M
Cañal
María Jesús
MJ
Rodríguez
Roberto
R
D000818:Animals; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000032:analysis; D018744:DNA, Plant / Q000032:analysis; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000592:standards; D016678:Genome; D006801:Humans
D004273:DNA, Neoplasm; D018744:DNA, Plant; D003841:Deoxycytidine; D004247:DNA; C016569:5-methyldeoxycytidine