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Determination of 5‐methyl‐cytosine and cytosine in tumor DNA of cancer patients
Author(s) -
Sandoval Guerrero Karina,
Revilla Vázquez Alma,
SeguraPacheco Blanca,
DueñasGonzalez Alfonso
Publication year - 2005
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/elps.200410040
Subject(s) - cytosine , guanine , capillary electrophoresis , thymine , chemistry , dna , uracil , chromatography , sodium dodecyl sulfate , methylation , dna methylation , analytical chemistry (journal) , biochemistry , gene , nucleotide , gene expression
Abstract The determination of the relative methylation in DNA tumor samples, in order to evaluate the activity of some anti‐cancer drugs, has become a very important issue in the clinical field. Capillary electrophoresis (CE) applications in this area have been done previously but no good separation for model samples or tumor samples has been reported. In this work, the CE conditions have been optimized in order to obtain baseline separation and efficient peaks for cytosine and 5‐methylcytosine in both, standard mixtures and actual tumor samples; other bases (adenine, uracil, guanine, and thymine) have also been integrated in the optimization studies. More efficient peaks and shorter analysis time compared with the already reported conditions have been obtained employing a fused‐silica capillary (75 µm inner diamter) of 44.5 cm effective length, 20 m M carbonate buffer (pH 9.6) plus 80 m M sodium dodecyl sulfate, a separation voltage of 20 kV, and detection at 223 nm.