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Normalized Multipotential Redox Coding of DNA Bases for Determination of Total Nucleotide Composition
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SYSNO ASEP 0574341 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Normalized Multipotential Redox Coding of DNA Bases for Determination of Total Nucleotide Composition Author(s) Kodr, David (UOCHB-X) ORCID
Ortiz, M. (ES)
Sýkorová, Veronika (UOCHB-X) ORCID, RID
Yenice, C. P. (ES)
Lesnikowski, Z. J. (PL)
O’Sullivan, C. K. (ES)
Hocek, Michal (UOCHB-X) RID, ORCIDSource Title Analytical Chemistry. - : American Chemical Society - ISSN 0003-2700
Roč. 95, č. 34 (2023), s. 12586-12589Number of pages 4 s. Language eng - English Country US - United States Keywords direct electrochemical detection ; nucleoside triphosphates ; aromatic groups OECD category Organic chemistry R&D Projects GA22-11252S GA ČR - Czech Science Foundation (CSF) GX20-00885X GA ČR - Czech Science Foundation (CSF) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 UT WOS 001050912800001 EID SCOPUS 85169028174 DOI 10.1021/acs.analchem.3c02023 Annotation The previously reported approach of orthogonal multipotential redox coding of all four DNA bases allowed only analysis of the relative nucleotide composition of short DNA stretches. Here, we present two methods for normalization of the electrochemical readout to facilitate the determination of the total nucleotide composition. The first method is based on the presence or absence of an internal standard of 7-deaza-2′-deoxyguanosine in a DNA primer. The exact composition of the DNA was elucidated upon two parallel analyses and the subtraction of the electrochemical signal intensities. The second approach took advantage of a 5′-viologen modified primer, with this fifth orthogonal redox label acting as a reference for signal normalization, thus allowing accurate electrochemical sequence analysis in a single read. Both approaches were tested using various sequences, and the voltammetric signals obtained were normalized using either the internal standard or the reference label and demonstrated to be in perfect agreement with the actual nucleotide composition, highlighting the potential for targeted DNA sequence analysis. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2024 Electronic address https://doi.org/10.1021/acs.analchem.3c02023
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