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Biology 2016, 5(4), 47; doi:10.3390/biology5040047

Optimization of Cyanine Dye Stability and Analysis of FRET Interaction on DNA Microarrays

Institute of Technical Chemistry, Leibniz University Hanover, Callinstr. 5, 30167 Hanover, Germany
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Academic Editor: Chris O’Callaghan
Received: 29 September 2016 / Revised: 10 November 2016 / Accepted: 24 November 2016 / Published: 30 November 2016
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Abstract

The application of DNA microarrays for high throughput analysis of genetic regulation is often limited by the fluorophores used as markers. The implementation of multi-scan techniques is limited by the fluorophores’ susceptibility to photobleaching when exposed to the scanner laser light. This paper presents combined mechanical and chemical strategies which enhance the photostability of cyanine 3 and cyanine 5 as part of solid state DNA microarrays. These strategies are based on scanning the microarrays while the hybridized DNA is still in an aqueous solution with the presence of a reductive/oxidative system (ROXS). Furthermore, the experimental setup allows for the analysis and eventual normalization of Förster-resonance-energy-transfer (FRET) interaction of cyanine-3/cyanine-5 dye combinations on the microarray. These findings constitute a step towards standardization of microarray experiments and analysis and may help to increase the comparability of microarray experiment results between labs. View Full-Text
Keywords: microarray; DNA; scanning; photobleaching; fluorophore; cyanine dye; FRET; ROXS; bioinformatics; bioanalytics microarray; DNA; scanning; photobleaching; fluorophore; cyanine dye; FRET; ROXS; bioinformatics; bioanalytics
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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von der Haar, M.; Heuer, C.; Pähler, M.; von der Haar, K.; Lindner, P.; Scheper, T.; Stahl, F. Optimization of Cyanine Dye Stability and Analysis of FRET Interaction on DNA Microarrays. Biology 2016, 5, 47.

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