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Interactions of fluorescent dye SYBR Green I with natural and 7-deazaguanine-modified DNA studied by fluorescence and electrochemical methods

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Abstract

SYBR Green I (SG) is a fluorescent dye applied in various techniques of DNA analysis, including fluorescent staining of electrophoretic gels, quantitative polymerase chain reaction, etc. SG binds selectively to double-stranded DNA via intercalation and minor groove interactions, resulting in a considerable enhancement of fluorescence of the dye. Modification of DNA by partial or full replacement of natural purine nucleobase guanine (G) with its synthetic analog 7-deazaguanine (G*) or its derivatives was shown to cause the SG fluorescence quenching. In this paper, we present a comparative study of interactions of SG with natural DNA fragments and with DNA fragments modified with G* by means of fluorescence and electrochemical methods. Competition between unmodified (forming strongly fluorescent complex with SG) and fully G*-modified (not contributing significantly to overall fluorescence signal) DNA fragments for the dye was studied via changes in the fluorescence intensity. In addition, association interactions of natural or G*-modified DNA with SG in solution were monitored by adsorptive transfer stripping square wave voltammetry at a pyrolytic graphite electrode using a signal of SG electrooxidation. We show that SG binds both natural and G*-modified DNA with similar apparent affinity and selectivity for the double-stranded DNA.

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Acknowledgments

This work was supported by the Czech Science Foundation (grant P206/11/1638 to M. F. and P206/12/2378 to L. H.) and by the ASCR (RVO 68081707).

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Authors and Affiliations

  1. Institute of Biophysics, Academy of Sciences of the Czech Republic, v.v.i., Kralovopolska 135, 612 65, Brno, Czech Republic

    Zdenka Dudová, Jan Špaček, Luděk Havran, Hana Pivoňková & Miroslav Fojta

  2. Central European Institute of Technology, Masaryk University, Kamenice 753/5, 625 00, Brno, Czech Republic

    Jan Špaček, Luděk Havran & Miroslav Fojta

Authors
  1. Zdenka Dudová
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  2. Jan Špaček
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  3. Luděk Havran
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  4. Hana Pivoňková
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  5. Miroslav Fojta
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Correspondence to Miroslav Fojta.

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Dudová, Z., Špaček, J., Havran, L. et al. Interactions of fluorescent dye SYBR Green I with natural and 7-deazaguanine-modified DNA studied by fluorescence and electrochemical methods. Monatsh Chem 147, 13–20 (2016). https://doi.org/10.1007/s00706-015-1578-5

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  • DOI: https://doi.org/10.1007/s00706-015-1578-5

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