Ms. Boram Kim Profile

Boram Kim

at Pusan National Univ

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Proceedings Article | 12 February 2011 Paper

DMSO effects on FRET to dye-labeled DNA in conjugated polymer-based DNA detection

Mijeong Kang, Boram Kim, Han Young Woo

Proceedings Volume 7908, 790809 (2011) https://doi.org/10.1117/12.874398

KEYWORDS: Polymers, Fluorescence resonance energy transfer, Optical properties, Absorption, Signal processing, Quantum efficiency, Sensors, Bromine, Signal detection, Luminescence

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Solvent effects were studied in fluorescence resonance energy transfer (FRET) from a cationic polyfluorene copolymer (FHQ, FPQ) to a fluorescein (Fl)-labeled oligonucleotide (ssDNA-Fl). Upon addition of dimethyl sulfoxide (DMSO), optical properties of the polymers and the probe dye were substantially modified. And the FRET-induced Fl emission was measured by directly exciting the polymer within the complex, polymer/ssDNA-Fl. The FRET signal was successfully modulated with changing the DMSO content. In the case of FHQ, the FRET-induced Fl emission was seriously quenched in phosphate buffer solution (PBS), while a salient FRET signal was observed in a 80 vol% DMSO/PBS mixture (36.8 time higher than that in PBS). The FPQ-sensitized FRET signal was also 3.8-fold amplified by the presence of DMSO. That result is from the decrease of hydrophobic interactions between the polymer and ssDNA-Fl, which induces the weaker polymer/ssDNA-Fl complexation with longer intermolecular separation. The gradual decrease in Fl PL quenching with increasing the DMSO content was investigated by measuring the Stern- Volmer quenching constants (3.3-4.2 × 10⁶ M^-1 in PBS, 0.56-1.1 x 10⁶ M^-1 in 80 vol% DMSO) in PBS/DMSO mixtures. The substantially reduced PL quenching would amplify the resulting FRET Fl signal. This approach suggests a simple way of modifying the fine-structure of polymer/ssDNA-Fl and improving the detection sensitivity in conjugated polymer-based FRET bioassays.

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