Premium
High‐performance capillary electrophoretic method for the quantification of 5‐methyl 2'‐deoxycytidine in genomic DNA: Application to plant, animal and human cancer tissues
Author(s) -
Fraga Mario F.,
Uriol Esther,
Diego L. Borja,
Berdasco María,
Esteller Manel,
Cañal María Jesús,
Rodríguez Roberto
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200206)23:11<1677::aid-elps1677>3.0.co;2-z
Subject(s) - capillary electrophoresis , dna , genomic dna , dna methylation , chemistry , chromatography , methylation , deoxycytidine , microbiology and biotechnology , electrophoresis , biochemistry , biology , cancer , gene , genetics , gene expression , gemcitabine
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.