Voltammetric determination of underivatized oligonucleotides on graphite electrodes based on their oxidation products.
A new electrochemical method to determine underivatized oligonucleotides is developed. The electro-oxidation of the adenine moieties of adsorbed oligonucleotides at elevated potentials on pyrolytic graphite electrodes (PGE) in neutral or alkaline media gives rise to electroactive products strongly adsorbed on the electrode surface. The extent of the redox processes of these products, with formal potential close to 0 V (vs Ag /AgCl) at pH 10, correlates well with the amount of parent oligonucleotide. Various electrochemical techniques have been compared and applied to the detection of specific DNA sequences and synthetic homopolynucleotides. Detection limits of 2 and 10 ng for (dA)20 and a 21-mer sequence of HIV-1, respectively, have been achieved using sample volumes of 10 microL. Moreover, the adsorbed oxidized oligonucleotide shows electrocatalytic activity toward the oxidation of NADH. The capability of the new method to detect DNA hybridization is discussed.
Departamento de Química Física y Analítica, Universidad de Oviedo, Spain.
Patricia
P
de-los-Santos-Alvarez
M Jesús
MJ
Lobo-Castañón
Arturo J
AJ
Miranda-Ordieres
Paulino
P
Tuñón-Blanco
D002692:Chlamydia trachomatis / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D004277:DNA, Single-Stranded; D004279:DNA, Viral / Q000032:analysis; D004566:Electrodes; D006108:Graphite; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D009841:Oligonucleotides / Q000032:analysis*; D010084:Oxidation-Reduction; D011061:Poly A / Q000032:analysis; D011066:Poly C / Q000032:analysis; D011071:Poly T / Q000032:analysis; D011067:Poly dA-dT / Q000032:analysis
D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D004279:DNA, Viral; D009841:Oligonucleotides; D011061:Poly A; D011071:Poly T; C015465:poly(dA); C023826:poly(dC); D011067:Poly dA-dT; D011066:Poly C; D006108:Graphite