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    • 5. 发明授权
    • Fluorescent quenching detection reagents and methods
    • 荧光猝灭检测试剂及方法
    • US06727356B1
    • 2004-04-27
    • US09457616
    • 1999-12-08
    • Michael W. ReedEugeny Alexander LukhtanovAlexander A. GallRobert O. Dempcy
    • Michael W. ReedEugeny Alexander LukhtanovAlexander A. GallRobert O. Dempcy
    • C07H1900
    • C07D519/00C07B2200/11C07C245/08C07F9/2408C07F9/572C07F9/65522C07F9/6561C07H21/00C09B29/0813C12Q1/6818C40B40/00C12Q2565/1025C12Q2521/319
    • Oligonucleotide-fluorophore-quencher conjugates wherein the fluorophore moiety 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-quencher-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 have substantially improved hybridization and signal to noise ratio properties.
    • 寡核苷酸 - 荧光团 - 猝灭剂缀合物,其中荧光团部分具有在约300至约800nm范围内的发射波长,或者猝灭剂包含取代的4-(苯基二氮烯基)苯胺结构的提供改善的信噪比和其它有利特征 杂交和相关测定。 寡核苷酸 - 荧光团 - 猝灭剂缀合物可以通过利用引入基于取代的4-(苯基二氮烯基)苯基胺结构的猝灭剂部分的新型亚磷酰胺试剂,和/或新型亚磷酰胺试剂合成,所述亚磷酰胺试剂掺入基于取代香豆素的荧光团部分,取代7 - 羟基-3H-吩恶嗪-3-酮或取代的5,10-二氢-10- [苯基]吡啶并[2,3-d; 6,5-d']二嘧啶-2,4,6,8-( 1H,3H,7H,9H,10H) - 四酮结构。 吡咯并[4,5-e]二氢吲哚-7-基}羰基)吡咯并[4,5-e]二氢吲哚-7-基]羰基}吡咯并[4,5- e]二氢吲哚-7-羧酸酯(DPI 3)部分作为次槽粘合剂和取代的4-(苯基二氮烯基)苯胺部分作为猝灭剂,并且具有显着改善的杂交和信噪比性质。