A Facile and Cost‐effective Electroanalytical Strategy for the Quantification of Deoxyguanosine and Deoxyadenosine in Oligonucleotides Using Screen‐printed Graphite Electrodes. Issue 12 (25th July 2016)

• Record Type: Journal Article
• Authors: Brotons, Ariadna; Sanjuán, Ignacio; Foster, Christopher W.; Banks, Craig E.; Vidal‐Iglesias, Francisco José; Solla‐Gullón, José; Iniesta, Jesús
• Is Part Of: Electroanalysis. Volume 28:Issue 12(2016)
• Page Start: 3066
• Page End: 3074
• Publication Date: 2016-07-25
• Subjects: Screen printed graphite electrode -- enzymatic digestion -- Square Wave Voltammetry -- oligonucleotides; Electrochemical analysis -- Periodicals; Chemistry, Analytical -- Periodicals; Electrochemistry -- Periodicals; 543.4
• DOI: 10.1002/elan.201600272
• Languages: English
• ISSNs: 1040-0397

Abstract

Abstract: The development of electroanalytical methods for the detection and quantification of nucleotides in DNA offers vital implications in assessing the degree of oxidation or epigenetic modification in DNA. Unfortunately, the electrochemical response of oligonucleotides is strongly influenced by the size, composition and nucleic base sequence. In this article, an optimized analytical procedure for the enzymatically breakdown of the oligonucleotides to their corresponding nucleotides for the evaluation of the electrochemical response through the use of square wave voltammetry (SWV) is presented. Enzymatic digestion of oligonucleotides has been optimized in terms of buffer composition, digestion time, strategy for stopping the enzymatic reaction and filtration requirement for enzyme removal, and then compared to an established protocol. Under the optimized protocol SWV response of a number of untreated and enzymatically digested six‐mer oligonucleotides, namely 5′‐GGGGGG‐3′, 5′‐AAAAAA‐3′, 5′‐CGCGCG‐3′ and 5′‐AAACGC‐3′ have been analysed, providing a higher sensitivity for the determination of guanosine and adenosine monophosphate species under digestion conditions with a more facile and cost effective procedure. The novel strategy for the enzymatically treated oligonucleotides in combination with the SWV response provides a proof of principle for feasible applications in the diagnosis of methylated guanosine in DNA as a potential biomarker due to its relation with cancer. Abstract :