Abstract
Conjugations of oligonucleotides, chromophores, and gold nanoparticles (GNPs) can be used for energy transfer assays to detect DNA. Herein, a homogenous Förster resonance energy transfer (FRET) system employing two-step modification of oligonucleotide on GNPs was reported. The distance between the donor (fluorescein attached onto DNA) and the acceptor (GNPs) was controlled by using the G-rich DNA. In the presence of porphyrin or berberine, which can act as ligands of G-quadruplexes, the G-rich DNA spacer can result into G-quadruplex structure. Therefore, the intimate contact between the fluorophore and the GNP results in efficient energy transfer and fluorescence quenching. After hybridization with target DNA, the G-quadruplex stretched and resulted in an enhancement of fluorescence. So the present FRET system can be used for target DNA sensing with detection limit as low as 40pM (S/N=3). In this study, a relation between the fluorescence quenching efficiency and GNP sizes was found and bigger GNPs had higher fluorescence enhancement after hybridization with target DNA.
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