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    Fluorescent quenching detection reagents and methods
    4.
    发明授权
    Fluorescent quenching detection reagents and methods 有权
    荧光猝灭检测试剂及方法

    公开(公告)号：US06727356B1

    公开(公告)日：2004-04-27

    申请号：US09457616

    申请日：1999-12-08

    申请人： Michael W. Reed ,  Eugeny Alexander Lukhtanov ,  Alexander A. Gall ,  Robert O. Dempcy

    发明人： Michael W. Reed ,  Eugeny Alexander Lukhtanov ,  Alexander A. Gall ,  Robert O. Dempcy

    IPC分类号： C07H1900

    CPC分类号： C07D519/00 ,  C07B2200/11 ,  C07C245/08 ,  C07F9/2408 ,  C07F9/572 ,  C07F9/65522 ,  C07F9/6561 ,  C07H21/00 ,  C09B29/0813 ,  C12Q1/6818 ,  C40B40/00 ,  C12Q2565/1025 ,  C12Q2521/319

    摘要： 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.

    摘要翻译： 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。

    Fluorescent quenching detection reagents and methods
    6.
    发明授权
    Fluorescent quenching detection reagents and methods 有权
    荧光猝灭检测试剂及方法

    公开(公告)号：US06699975B2

    公开(公告)日：2004-03-02

    申请号：US10093769

    申请日：2002-03-07

    申请人： Michael W. Reed ,  Eugeny Alexander Lukhtanov ,  Alexander A. Gall ,  Robert O. Dempcy

    发明人： Michael W. Reed ,  Eugeny Alexander Lukhtanov ,  Alexander A. Gall ,  Robert O. Dempcy

    IPC分类号： C07C24500

    CPC分类号： C07D519/00 ,  C07B2200/11 ,  C07C245/08 ,  C07F9/2408 ,  C07F9/572 ,  C07F9/65522 ,  C07F9/6561 ,  C07H21/00 ,  C09B29/0813 ,  C12Q1/6818 ,  C40B40/00 ,  C12Q2565/1025 ,  C12Q2521/319

    摘要： 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-(phenyidiazenyl)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.

    摘要翻译： 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，或者其中猝灭剂包括取代的4-（苯基二氮烯基）苯胺结构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基于取代的4-（苯基亚基）苯胺结构的掺入猝灭剂部分的新型亚磷酰胺试剂和/或掺入基于取代香豆素的荧光团部分的新型亚磷酰胺试剂来合成，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。

    Attachment of oligonucleotides to solid supports through Schiff base type linkages for capture and detection of nucleic acids
    7.
    发明授权
    Attachment of oligonucleotides to solid supports through Schiff base type linkages for capture and detection of nucleic acids 失效
    通过席夫碱型连接将寡核苷酸与固体支持物连接，用于捕获和检测核酸

    公开(公告)号：US06339147B1

    公开(公告)日：2002-01-15

    申请号：US09364320

    申请日：1999-07-29

    申请人： Eugeny Alexander Lukhtanov ,  Mikhail A. Podyminogin ,  Joel Hedgpeth

    发明人： Eugeny Alexander Lukhtanov ,  Mikhail A. Podyminogin ,  Joel Hedgpeth

    IPC分类号： C07H2100

    CPC分类号： C40B50/18 ,  C07B2200/11 ,  C07F9/2408 ,  C07H21/00

    摘要： Derivatized oligonucleotides (ODNs) are coupled to a solid support in improved yield resulting in a high density of coupled oligonucleotide per surface unit of the support, through a Schiff base type bond formed between an NH2 group attached either to the solid support or to the ODN and an aromatic aldehyde attached to the other of the solid support and the ODN. The preferred solid support-ODN conjugate is formed between semicarbazide groups attached to a glass surface and an aromatic aldehyde attached at either 3′, or 5′ end of an ODN or to an intermediate nucleotide of the ODN.

    摘要翻译： 衍生化的寡核苷酸（ODN）以提高的产量与固体支持物偶联，通过在连接到固体支持物上的NH 2基团或与ODN之间形成的席夫碱型键，形成每个载体每个表面单元的高密度耦合寡核苷酸 和连接到另一个固体支持物和ODN的芳族醛。 优选的固体支持物-ODN缀合物在连接到玻璃表面的氨基脲基团和连接在ODN的3'或5'末端的芳香醛或ODN的中间核苷酸之间形成。

    Attachment of oligonucleotides to solid supports through Schiff base type linkages for capture and detection of nucleic acids
    9.
    发明授权
    Attachment of oligonucleotides to solid supports through Schiff base type linkages for capture and detection of nucleic acids 有权
    通过席夫碱型连接将寡核苷酸与固体支持物连接，用于捕获和检测核酸

    公开(公告)号：US06441159B1

    公开(公告)日：2002-08-27

    申请号：US09780908

    申请日：2001-02-09

    申请人： Eugeny Alexander Lukhtanov ,  Mikhail A. Podyminogin

    发明人： Eugeny Alexander Lukhtanov ,  Mikhail A. Podyminogin

    IPC分类号： C07H2100

    CPC分类号： C40B50/18 ,  C07B2200/11 ,  C07F9/2408 ,  C07H21/00

    摘要： Derivatized oligonucleotides (ODNs) are coupled to a solid support in improved yield resulting in a high density of coupled oligonucleotide per surface unit of the support through a Schiff base type bond formed between an NH2 group attached either to the solid support or to the ODN and an aromatic aldehyde attached to the other of the solid support and the ODN. The preferred solid support-ODN conjugate is formed between semicarbazide groups attached to a glass surface and an aromatic aldehyde attached at either 3′, or 5′ end of an ODN or to an intermediate nucleotide of the ODN.

    摘要翻译： 衍生的寡核苷酸（ODN）以提高的产率偶联到固体支持物上，通过在连接到固体支持物上的NH 2基团或与ODN之间形成的席夫碱型键，形成每个表面单位的载体的高密度偶联的寡核苷酸，以及 连接到另一个固体支持物和ODN的芳族醛。 优选的固体支持物-ODN缀合物在连接到玻璃表面的氨基脲基团和连接在ODN的3'或5'末端的芳香醛或ODN的中间核苷酸之间形成。