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公开(公告)号:US20030148470A1
公开(公告)日:2003-08-07
申请号:US10192776
申请日:2002-07-09
申请人: PE Corporation (NY)
IPC分类号: C12Q001/68 , C12P019/34 , C07H021/04 , C07H019/067 , C07H019/04 , C07H019/16 , C07H019/06 , C07H019/073 , C07H019/09
摘要: Propargylethoxyamino nucleosides are disclosed having the structure 1 wherein R1 and R2 are nullH, lower alkyl, or label; B is a 7-deazapurine, purine, or pyrimidine nucleoside base; W1 is nullH or nullOH; W2 is nullOH or a moiety which renders the nucleoside incapable of forming a phosphodiester bond at the 3null-position; and W3 is nullPO4, nullP2O7, nullP3O10, phosphate analog, or nullOH. Additionaly, a primer extension method is provided employing the above propargylethoxyamino nucleosides.
摘要翻译: 公开了具有其中R 1和R 2为-H,低级烷基或标记的结构的丙炔乙氧基氨基核苷; B是7-脱氮嘌呤,嘌呤或嘧啶核苷碱基; W1是-H或-OH; W2是-OH或使核苷不能在3'-位形成磷酸二酯键的部分; W3是-PO 4,-P 2 O 7,-P 3 O 10,磷酸酯类似物或-OH。 另外,使用上述炔丙氧基氨基核苷提供引物延伸方法
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公开(公告)号:US20010014735A1
公开(公告)日:2001-08-16
申请号:US09813378
申请日:2001-03-20
申请人: PE Corporation (NY)
IPC分类号: C07H021/04 , C07H021/02 , C07H021/00
摘要: Methods and compositions to label oligonucleotides and analogs directly on a solid-support having the structure 1 where S is a solid-support, A is a cleavable linker, X is a moiety with three or more attachment sites, L is a label, Y is a nucleophile, i.e. O, NH, NR or S, and P1 is an acid cleavable protecting group are provided. The labelled solid-support is reacted in a cyclical fashion to synthesize a labelled oligonucleotide on a solid-support in the 5null to 3null direction, having the structure: 2 Labelled oligonucleotides are also synthesized by reacting: (i) a label reagent bearing functionality consisting of carboxylic acid, sulfonic acid, phosphonic acid, or phosphoric acid, (ii) an oligonucleotide on solid support with nucleophilic functionality, and (iii) a coupling reagent, whereby an ester, amide, thioester, sulfonamide, sulfonate, phosphonate, phosphoramidate, phosphorothioate, or phosphate bond is formed. The labelling reaction may be conducted at label sites including the 5null terminus, the 3null terminus, a nucleobase, an internucleotide linkage, a sugar, amino, sulfide, hydroxyl, and carboxyl.
摘要翻译: 方法和组合物直接在具有S是固体支持物的结构的固体支持物上标记寡核苷酸和类似物,A是可切割的连接体,X是具有三个或更多个连接位点的部分,L是标记,Y是一个 亲核试剂,即O,NH,NR或S,P1是可酸切割的保护基团。 标记的固体支持物以循环方式反应以在5'至3'方向的固体支持物上合成标记的寡核苷酸,具有以下结构:标记的寡核苷酸还通过以下方式合成:(i)标记试剂载体官能团 包括羧酸,磺酸,膦酸或磷酸,(ii)具有亲核官能团的固体支持物上的寡核苷酸,和(iii)偶联剂,其中酯,酰胺,硫酯,磺酰胺,磺酸酯,膦酸酯,氨基磷酸酯 ,硫代磷酸酯或磷酸酯键。 标记反应可以在包括5'末端,3'末端,核碱基,核苷酸间键,糖,氨基,硫化物,羟基和羧基的标记位点进行。
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公开(公告)号:US20020151711A1
公开(公告)日:2002-10-17
申请号:US10085561
申请日:2002-02-26
申请人: PE Corporation (NY)
IPC分类号: C07H021/04 , C12Q001/68
CPC分类号: C07H21/00 , C07H19/04 , C07H19/06 , C07H19/14 , C07H19/16 , C07H19/20 , C12Q1/6869 , C12Q2525/186 , C12Q2525/117 , C12Q2525/113
摘要: A nucleoside/tide compound having the structure NUCnullLnullSnullLB/LG is described wherein NUC is a nucleoside/tide having a nucleobase portion B, L is a rigid linkage, S is a spacer; and LB/LG is a member of a linkage pair or a label. NUC is attached to L through B such that when B is a purine, L is attached to the 8-position of the purine, when B is 7-deazapurine, L is attached to the 7-position of the 7-deazapurine, and when B is pyrimidine, L is attached to the 5-position of the pyrimidine. In an important aspect of the invention, L has the structure 1 wherein each of n, o and p are integers ranging from 0 to 3, and the sum of n, o and p is at least 2, and each of W, X, Y and Z is selected from the group consisting of carbon and nitrogen. The invention further includes polynucleotide compounds comprising the nucleoside/tide, and primer extension methods utilizing the nucleoside/tide, particularly when used in combination with certain mutant polymerase enzymes.
摘要翻译: 其中NUC是具有核碱基的核苷/潮汐的核苷/潮汐化合物,其具有结构<片段-inline-formula> NUC-LS-LB / LG in-line-formula> 部分B,L是刚性连接,S是间隔物; 而LB / LG是连锁对或标签的成员。 N与B连接,B为嘌呤时,L为嘌呤8位时,B为7-脱氮嘌呤时,L为7-脱氮嘌呤的7-位, B是嘧啶,L连接到嘧啶的5位上。 在本发明的一个重要方面,L具有其中n,o和p各自为0〜3的整数,n,o和p的和为2以上的结构,W,X,Y Z选自碳和氮。 本发明还包括包含核苷/潮汐的多核苷酸化合物,以及利用核苷/潮汐的引物延伸方法,特别是当与某些突变型聚合酶组合使用时。
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4.
公开(公告)号:US20020165389A1
公开(公告)日:2002-11-07
申请号:US10010717
申请日:2001-11-07
申请人: PE Corporation (NY)
IPC分类号: C07H021/04
摘要: Methods and compositions to label oligonucleotides and analogs directly on a solid-support having the structure 1 where S is a solid-support, A is a cleavable linker, X is a moiety with three or more attachment sites, L is a label, Y is a nucleophile, i.e. O, NH, NR or S, and P1 is an acid cleavable protecting group are provided. The labelled solid-support is reacted in a cyclical fashion to synthesize a labelled oligonucleotide on a solid-support in the 5null to 3null direction, having the structure: 2 Labelled oligonucleotides are also synthesized by reacting: (i) a label reagent bearing functionality consisting of carboxylic acid, sulfonic acid, phosphonic acid, or phosphoric acid, (ii) an oligonucleotide on solid support with nucleophilic functionality, and (iii) a coupling reagent, whereby an ester, amide, thioester, sulfonamide, sulfonate, phosphonate, phosphoramidate, phosphorothioate, or phosphate bond is formed. The labelling reaction may be conducted at label sites including the 5null terminus, the 3null terminus, a nucleobase, an internucleotide linkage, a sugar, amino, sulfide, hydroxyl, and carboxyl.
摘要翻译: 方法和组合物直接在具有S是固体支持物的结构的固体支持物上标记寡核苷酸和类似物,A是可切割的连接体,X是具有三个或更多个连接位点的部分,L是标记,Y是一个 亲核试剂,即O,NH,NR或S,P1是可酸切割的保护基团。 标记的固体支持物以循环方式反应以在5'至3'方向的固体支持物上合成标记的寡核苷酸,具有以下结构:标记的寡核苷酸还通过以下方式合成:(i)标记试剂载体官能团 包括羧酸,磺酸,膦酸或磷酸,(ii)具有亲核官能团的固体支持物上的寡核苷酸,和(iii)偶联试剂,其中酯,酰胺,硫酯,磺酰胺,磺酸酯,膦酸酯,氨基磷酸酯 ,硫代磷酸酯或磷酸酯键。 标记反应可以在包括5'末端,3'末端,核碱基,核苷酸间键,糖,氨基,硫化物,羟基和羧基的标记位点进行。
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公开(公告)号:US20020102590A1
公开(公告)日:2002-08-01
申请号:US09976168
申请日:2001-10-11
申请人: PE Corporation (NY)
IPC分类号: C12Q001/68 , C07H019/04 , C07D487/22 , C07D473/00
摘要: Provided are nucleotide-dye conjugates and related compounds in which a dye is linked to a nucleobase directly or indirectly by an anionic linker. The anionic character of the linker is provided by one or more anionic moieties which are present in the linker, such as phosphate, phosphonate, sulfonate, and carboxylate groups. When the dye is a provided as a donor/acceptor dye pair, the anionic linker can be located between the donor and the acceptor, or between the nucleobase and either the donor or acceptor, or both. In one embodiment, conjugates of the invention provide enhanced electrophoretic mobility characteristics to sequencing fragments, e.g., for dideoxy sequencing using labeled terminators.
摘要翻译: 提供的是核苷酸 - 染料缀合物和相关化合物,其中染料通过阴离子连接体直接或间接连接到核碱基。 连接体的阴离子特征由连接物中存在的一个或多个阴离子部分提供,例如磷酸酯,膦酸酯,磺酸酯和羧酸酯基团。 当染料作为供体/受体染料对提供时,阴离子连接体可以位于供体和受体之间,或位于核碱基与供体或受体之间或两者之间。 在一个实施方案中,本发明的缀合物为测序片段提供增强的电泳迁移率特性,例如使用标记的终止子进行双脱氧测序。
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公开(公告)号:US20030055243A1
公开(公告)日:2003-03-20
申请号:US10227058
申请日:2002-08-21
申请人: PE Corporation (NY)
IPC分类号: C09B047/04 , C09B062/00 , C09B067/00
摘要: Atropisomeric energy-transfer dye compounds are disclosed. A variety of molecular biology applications utilize atropisomeric xanthene fluorescent dyes as labels for substrates such as nucleotides, nucleosides, polynucleotides, polypeptides and carbohydrates. Methods include DNA sequencing, DNA fragment analysis, PCR, SNP analysis, oligonucleotide ligation, amplification, minisequencing, and primer extension.
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