Electrochemical determination of d(GCGAAAGC) hairpin

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Authors

TRNKOVÁ Libuše POSTBIEGLOVÁ Irena HOLÍK Miroslav

Year of publication 2003
Type Article in Proceedings
Conference XVII th International Symposium on Bioelectrochemistry and Bioenergetics
MU Faculty or unit

Faculty of Science

Citation
Field Electrochemistry
Keywords hairpin; linear sweep and cyclic voltammetry; elimination voltammetry with linear scan; Fourier Transform;confidence ellipse
Description Hairpins (or hairpin-like structures) may play a major role in expansion events of triplet repeat expansion diseases (X syndrome, Huntington s disease, Friedeich s ataxia). The d(GCGAAGC) fragment has been found in the replication origins of phage fX 174 and herpes simplex virus, in a promoter region of an E.coli heat-shock gene, and in rRNA genes. Recently, the structure of d(GCGAAGC) has been determined by NMR spectroscopy and refined using molecular dynamics [1]. For fast and economical determination of the hairpin in biological materials applicability of voltammetric methods was investigated. On mercury electrodes the d(GCGAAGC) hairpin provides a voltammetric reduction signal (A and C) and an oxidation signal (G). Both signals have been studied by cyclic voltammetry and square wave voltammetry in dependence on pH change, accumulation time, scan rate, and exchange of minihairpin sequences. The elimination voltammetry with linear scan (EVLS) [2-4] in combination with the adsorptive stripping the A and C reduction peak in the d(GCGXAGC) hairpin (X = A,C,T,G) has been employed to the determination of detection limits (~ nM). Multidimensional data of CV and EVLS were worked up by Fourier Transform (FT) and for the first coefficient a confidence ellipse was calculated in order to drop out some outlier data. The same method was used for detection limit determinations.
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