Quencher-free linear probe with multiple fluorophores on an acyclic scaffold

Abstract

We have developed a new quencher-free stemless linear probe involving multiple perylenes incorporated through D-threoninol; each perylene is separated by intervening natural nucleotides. Without a substrate, the flexible linear probe does not emit fluorescence due to the self-quenching of the weakly interacting fluorophores. Upon hybridization with the target, intercalation of each dye between the base pairs results in emission of strong fluorescence. The maximum signal–background ratio attained was 180, and the response rate was significantly faster than that of a classic hairpin-forming molecular beacon.