| 摘要 |
Oligonucleotide-fluorophore-quencher conjugates wherein the fluorophore moie ty has emission wavelengths in the range of about (300) to about (800) nm, and or where the quencher includes a substituted 4-(phenyldiazenyl)phenylamine structure provide improved signal to noise ratios and other advantageous characteristics in hybridization and related assays. The oligonucleotide- fluorophore-quencher conjugates can be synthesized by utilizing novel phosphoramidite reagents that incorporate the quencher moiety based on the substituted 4-(phenyldiazenyl)phenylamine structure, and or novel phosphoramidite reagents that incorporate a fluorophore moiety based on the substituted coumarin, substituted 7-hydroxy-3H-phenoxazin-3-one, or substituted 5,10-dihydro-10-[phenyl]pyrido[2,3-d;6,5-d']dipyrimidine-2,4,6,8 - (1H, 3H, 7H, 9H, 10H)-tetrone structure. Oligonucleotide-fluorophore-quenche r- minor groove binder conjugates including a pyrrolo[4,5-e]indolin-7- yl}carbonyl)pyrrolo[4,5-e]indolin-7-yl]carbonyl}pyrrolo[4,5-e]indoline-7- carboxylate (DPI3) moiety as the minor groove binder and the substituted 4- (phenyldiazenyl)phenylamine moiety as the quencher, were synthesized and hav e substantially improved hybridization and signal to noise ratio properties.</ SDOAB>
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