CTAB enhancement of FRET in DNA structures
The effect of cetyl‐trimethylammonium bromide (CTAB) on enhancing the fluorescence resonance energy transfer (FRET) between two dye‐conjugated DNA strands was studied using fluorescence emission spectroscopy and dynamic light scattering (DLS). For hybridized DNA where one strand is conjugated with a...
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Veröffentlicht in: | Journal of biophotonics 2016-01, Vol.9 (1-2), p.49-54 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | The effect of cetyl‐trimethylammonium bromide (CTAB) on enhancing the fluorescence resonance energy transfer (FRET) between two dye‐conjugated DNA strands was studied using fluorescence emission spectroscopy and dynamic light scattering (DLS). For hybridized DNA where one strand is conjugated with a TAMRA donor and the other with a TexasRed acceptor, increasing the concentration of CTAB changes the fluorescence emission properties and improves the FRET transfer efficiency through changes in the polarity of the solvent, neutralization of the DNA backbone and micelle formation. For the DNA FRET system without CTAB, the DNA hybridization leads to contact quenching between TAMRA donor and TexasRed acceptor producing reduced donor emission and only a small increase in acceptor emission. At 50 µM CTAB, however, the sheathing and neutralization of the dye‐conjugated dsDNA structure significantly reduces quenching by DNA bases and dye interactions, producing a large increase in FRET efficiency, which is almost four fold higher than without CTAB.
Adding CTAB to the aqueous buffer solution not only neutralized DNA strand, but also the forming DNA‐CTAB complex, which affected the fluorescence emissions of the dyes on the DNA. When CTAB molecules surround DNA strand and sheath/shield/insulate the donor (TAMRA) and acceptor (TexasRed) on the DNA, FRET enhanced due to reduction of the contact quenching by CTAB and closer proximity of two dyes. |
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ISSN: | 1864-063X 1864-0648 |
DOI: | 10.1002/jbio.201500221 |