公开(公告)号：US07541444B2

公开(公告)日：2009-06-02

申请号：US10525401

申请日：2003-08-22

申请人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

发明人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

IPC分类号： C07G11/00 ,  C07H21/02 ,  C12Q1/68 ,  C12P19/34 ,  C07H21/00

CPC分类号： C12Q1/6869 ,  C07H1/00 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C12Q2525/186 ,  C12Q2535/113 ,  Y02P20/55 ,  C12Q2525/197

摘要： The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O-Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R′)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula ═C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H.

公开(公告)号：US08071739B2

公开(公告)日：2011-12-06

申请号：US12804352

申请日：2010-07-20

申请人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

发明人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

IPC分类号： C07H21/02 ,  C07H21/04 ,  C07H19/04 ,  C12Q1/68

CPC分类号： C12Q1/6869 ,  C07H1/00 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C12Q2525/186 ,  C12Q2535/113 ,  Y02P20/55 ,  C12Q2525/197

摘要： The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O—Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R′)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula ═C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H.

摘要翻译： 本发明提供了修饰的核苷酸或核苷分子，其包含嘌呤或嘧啶碱基和核糖或脱氧核糖糖部分，其具有共价连接到其上的可移除的3'-OH封闭基团，使得3'碳原子连接一组结构-O -Z其中Z是-C（R'）2 -O-R“，-C（R'）2 -N（R”）2，-C（R'）2 -N（H）R“ -C（R'）2 -S-R“和-C（R'）2 -F，其中每个R”为或可移除的保护基团的一部分; 每个R'独立地是氢原子，烷基，取代的烷基，芳基烷基，烯基，炔基，芳基，杂芳基，杂环，酰基，氰基，烷氧基，芳氧基，杂芳氧基或酰氨基，或通过连接基团连接的可检测标记; 或（R'）2表示式≡C（R'“）2的亚烷基，其中每个R”可以相同或不同，并且选自氢和卤素原子和烷基; 并且其中所述分子可以反应以产生其中每个R“被交换为H的中间体，或其中Z是-C（R'）2 -F，F被交换成OH，SH或NH 2，优选OH，其中 中间体在水性条件下解离得到具有游离3'OH的分子; 条件是其中Z是-C（R'）2 -S-R“，两个R'基团不是H.

公开(公告)号：US20070166705A1

公开(公告)日：2007-07-19

申请号：US10525401

申请日：2003-08-22

申请人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

发明人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12P19/34 ,  C07F9/6512

CPC分类号： C12Q1/6869 ,  C07H1/00 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C12Q2525/186 ,  C12Q2535/113 ,  Y02P20/55 ,  C12Q2525/197

摘要： The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O-Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R′)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula ═C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H.

摘要翻译： 本发明提供了修饰的核苷酸或核苷分子，其包含嘌呤或嘧啶碱基和核糖或脱氧核糖糖部分，其具有共价连接到其上的可移除的3'-OH封闭基团，使得3'碳原子连接有一组结构-OZ 其中Z是-C（R'）2 -O-R“，-C（R'）2 -N（R”）2，-C（R'）2 -N（H）R“ ，-C（R'）2 -S-R“和-C（R'）2 -F，其中每个R”或是可除去的保护基团的一部分; 每个R'独立地是氢原子，烷基，取代的烷基，芳基烷基，烯基，炔基，芳基，杂芳基，杂环，酰基，氰基，烷氧基，芳氧基，杂芳氧基或酰氨基，或通过连接基团连接的可检测标记; 或（R'）2表示式-C（R“'）2的亚烷基，其中每个R”可以相同或不同，并且选自氢和卤素原子和烷基; 并且其中所述分子可以反应以产生其中每个R“被交换为H的中间体，或者其中Z是-C（R'）2-F，F被交换成OH，SH或NH 2，优选OH， 该中间体在水性条件下解离以得到具有游离3'OH的分子; 条件是其中Z是-C（R'）2 -S-R“，两个R'基团都不是H.

公开(公告)号：US08597881B2

公开(公告)日：2013-12-03

申请号：US13281275

申请日：2011-10-25

申请人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

发明人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

IPC分类号： C12Q1/68 ,  C07H19/04

CPC分类号： C12Q1/6869 ,  C07H1/00 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C12Q2525/186 ,  C12Q2535/113 ,  Y02P20/55 ,  C12Q2525/197

摘要： Provided herein is a method for replacing the allyl group in a compound of formula R—O-allyl, R2N(allyl), RNH(allyl), RN(allyl)2 or R—S-allyl with a hydrogen, using a transition metal and one or more phosphine ligands.

摘要翻译： 本文提供的是使用过渡金属和一种或多种式（I）的式RO-烯丙基，R2N（烯丙基），RNH（烯丙基），RN（烯丙基）2或RS-烯丙基的化合物中的烯丙基 膦配体。

公开(公告)号：US07771973B2

公开(公告)日：2010-08-10

申请号：US12455397

申请日：2009-06-01

申请人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

发明人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

IPC分类号： C12P19/34 ,  C12Q1/68 ,  C07H21/02 ,  C07H21/04

CPC分类号： C12Q1/6869 ,  C07H1/00 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C12Q2525/186 ,  C12Q2535/113 ,  Y02P20/55 ,  C12Q2525/197

摘要： The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O—Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R″)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula ═C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H.

摘要翻译： 本发明提供了修饰的核苷酸或核苷分子，其包含嘌呤或嘧啶碱基和核糖或脱氧核糖糖部分，其具有共价连接到其上的可移除的3'-OH封闭基团，使得3'碳原子连接一组结构-O -Z其中Z是-C（R'）2 -O-R“，-C（R'）2 -N（R”）2，-C（R'）2 -N（H）R“ -C（R'）2 -S-R“和-C（R”）2 -F，其中每个R“为或可除去保护基团的一部分; 每个R'独立地是氢原子，烷基，取代的烷基，芳基烷基，烯基，炔基，芳基，杂芳基，杂环，酰基，氰基，烷氧基，芳氧基，杂芳氧基或酰氨基，或通过连接基团连接的可检测标记; 或（R'）2表示式≡C（R'“）2的亚烷基，其中每个R”可以相同或不同，并且选自氢和卤素原子和烷基; 并且其中所述分子可以反应以产生其中每个R“被交换为H的中间体，或其中Z是-C（R'）2 -F，F被交换成OH，SH或NH 2，优选OH，其中 中间体在水性条件下解离得到具有游离3'OH的分子; 条件是其中Z是-C（R'）2 -S-R“，两个R'基团不是H.

公开(公告)号：US20120095201A1

公开(公告)日：2012-04-19

申请号：US13281275

申请日：2011-10-25

申请人： John MILTON ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

发明人： John MILTON ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

IPC分类号： C07H1/00

CPC分类号： C12Q1/6869 ,  C07H1/00 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C12Q2525/186 ,  C12Q2535/113 ,  Y02P20/55 ,  C12Q2525/197

摘要： Provided herein is a method for replacing the allyl group in a compound of formula R—O-allyl, R2N(allyl), RNH(allyl), RN(allyl)2 or R—S-allyl with a hydrogen, using a transition metal and one or more phosphine ligands.

摘要翻译： 本文提供了使用过渡金属替代式R-O-烯丙基，R2N（烯丙基），RNH（烯丙基），RN（烯丙基）2或R-S-烯丙基的烯丙基的方法 和一种或多种膦配体。

公开(公告)号：US20100292452A1

公开(公告)日：2010-11-18

申请号：US12804352

申请日：2010-07-20

申请人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

发明人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

IPC分类号： C07H21/00 ,  C07H21/04

CPC分类号： C12Q1/6869 ,  C07H1/00 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C12Q2525/186 ,  C12Q2535/113 ,  Y02P20/55 ,  C12Q2525/197

摘要： The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O—Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R′)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula ═C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H.

摘要翻译： 本发明提供了修饰的核苷酸或核苷分子，其包含嘌呤或嘧啶碱基和核糖或脱氧核糖糖部分，其具有共价连接到其上的可移除的3'-OH封闭基团，使得3'碳原子连接一组结构-O -Z其中Z是-C（R'）2 -O-R“，-C（R'）2 -N（R”）2，-C（R'）2 -N（H）R“ -C（R'）2 -S-R“和-C（R'）2 -F，其中每个R”为或可移除的保护基团的一部分; 每个R'独立地是氢原子，烷基，取代的烷基，芳基烷基，烯基，炔基，芳基，杂芳基，杂环，酰基，氰基，烷氧基，芳氧基，杂芳氧基或酰氨基，或通过连接基团连接的可检测标记; 或（R'）2表示式≡C（R'“）2的亚烷基，其中每个R”可以相同或不同，并且选自氢和卤素原子和烷基; 并且其中所述分子可以反应以产生其中每个R“被交换为H的中间体，或其中Z是-C（R'）2 -F，F被交换成OH，SH或NH 2，优选OH，其中 中间体在水性条件下解离得到具有游离3'OH的分子; 条件是其中Z是-C（R'）2 -S-R“，两个R'基团不是H.

公开(公告)号：US20090325172A1

公开(公告)日：2009-12-31

申请号：US12455397

申请日：2009-06-01

申请人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

发明人： John Milton ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Barnes ,  Xiaohai Liu ,  Silke Ruediger

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6869 ,  C07H1/00 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C12Q2525/186 ,  C12Q2535/113 ,  Y02P20/55 ,  C12Q2525/197

摘要： The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O-Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R′)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula ═C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H.

摘要翻译： 本发明提供了修饰的核苷酸或核苷分子，其包含嘌呤或嘧啶碱基和核糖或脱氧核糖糖部分，其具有共价连接到其上的可移除的3'-OH封闭基团，使得3'碳原子连接有一组结构-OZ 其中Z是-C（R'）2 -O-R“，-C（R'）2 -N（R”）2，-C（R'）2 -N（H）R“ ，-C（R'）2 -S-R“和-C（R'）2 -F，其中每个R”或是可除去的保护基团的一部分; 每个R'独立地是氢原子，烷基，取代的烷基，芳基烷基，烯基，炔基，芳基，杂芳基，杂环，酰基，氰基，烷氧基，芳氧基，杂芳氧基或酰氨基，或通过连接基团连接的可检测标记; 或（R'）2表示式-C（R“'）2的亚烷基，其中每个R”可以相同或不同，并且选自氢和卤素原子和烷基; 并且其中所述分子可以反应以产生其中每个R“被交换为H的中间体，或者其中Z是-C（R'）2-F，F被交换成OH，SH或NH 2，优选OH， 该中间体在水性条件下解离以得到具有游离3'OH的分子; 条件是其中Z是-C（R'）2 -S-R“，两个R'基团都不是H.

公开(公告)号：US20130197209A1

公开(公告)日：2013-08-01

申请号：US13791575

申请日：2013-03-08

申请人： John MILTON ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Bames ,  Xiaohai Liu ,  Silke Ruediger

发明人： John MILTON ,  Xiaolin Wu ,  Mark Smith ,  Joseph Brennan ,  Colin Bames ,  Xiaohai Liu ,  Silke Ruediger

IPC分类号： C07H1/00

CPC分类号： C12Q1/6869 ,  C07H1/00 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C12Q2525/186 ,  C12Q2535/113 ,  Y02P20/55 ,  C12Q2525/197

摘要： The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3′-OH blocking group covalently attached thereto, such that the 3′ carbon atom has attached a group of the structure —O—Z wherein Z is any of —C(R′)2-O—R″, —C(R′)2-N(R″)2, —C(R′)2-N(H)R″, —C(R′)2-S—R″ and —C(R′)2-F, wherein each R″ is or is part of a removable protecting group; each R′ is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R′)2 represents an alkylidene group of formula ═C(R′″)2 wherein each R′″ may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R″ is exchanged for H or, where Z is —C(R′) 2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3′OH; with the proviso that where Z is —C(R′)2-S—R″, both R′ groups are not H.

摘要翻译： 本发明提供了修饰的核苷酸或核苷分子，其包含嘌呤或嘧啶碱基和核糖或脱氧核糖糖部分，其具有共价连接到其上的可移除的3'-OH封闭基团，使得3'碳原子连接有一组结构-OZ 其中Z是-C（R'）2 -O-R“，-C（R'）2 -N（R”）2，-C（R'）2 -N（H）R“ ，-C（R'）2 -S-R“和-C（R'）2 -F，其中每个R”或是可除去的保护基团的一部分; 每个R'独立地是氢原子，烷基，取代的烷基，芳基烷基，烯基，炔基，芳基，杂芳基，杂环，酰基，氰基，烷氧基，芳氧基，杂芳氧基或酰氨基，或通过连接基团连接的可检测标记; 或（R'）2表示式= C（R“'）2的亚烷基，其中每个R”可以相同或不同，并且选自氢和卤素原子和烷基; 并且其中所述分子可以反应以产生其中每个R“被交换为H的中间体，或者其中Z是-C（R'）2-F，F被交换成OH，SH或NH 2，优选OH， 该中间体在水性条件下解离以得到具有游离3'OH的分子; 条件是其中Z是-C（R'）2 -S-R“，两个R'基团都不是H.

公开(公告)号：US20090118128A1

公开(公告)日：2009-05-07

申请号：US11989169

申请日：2006-07-20

申请人： Xiaohai Liu ,  John Milton ,  Geoffrey Paul Smith ,  Colin Barnes ,  Isabelle Marie Julia Rasolonjatovo ,  Roberto Rigatti ,  Xiaolin Wu ,  Tobias William Barr Ost ,  Graham John Worsley ,  David James Earnshaw ,  Gerardo Turcatti ,  Anthony Romieu

发明人： Xiaohai Liu ,  John Milton ,  Geoffrey Paul Smith ,  Colin Barnes ,  Isabelle Marie Julia Rasolonjatovo ,  Roberto Rigatti ,  Xiaolin Wu ,  Tobias William Barr Ost ,  Graham John Worsley ,  David James Earnshaw ,  Gerardo Turcatti ,  Anthony Romieu

IPC分类号： C40B20/00 ,  C07H1/00 ,  C12P19/34 ,  C40B50/06

CPC分类号： B01J19/0046 ,  B01J2219/00596 ,  B01J2219/00608 ,  B01J2219/00675 ,  B01J2219/00722 ,  C12Q1/6806 ,  C12Q1/6834 ,  C12Q1/6874 ,  C12Q2565/543 ,  C12Q2535/101 ,  C12Q2523/113 ,  C12Q2523/107 ,  C12Q2521/331 ,  C12Q2521/307

摘要： The invention relates to methods of generating templates for a nucleic acid sequencing reaction which comprise: providing at least one double-stranded nucleic acid molecule, wherein both strands of the double-stranded nucleic acid molecule are attached to a solid support at the 5′ end, cleaving one or both strands of the double-stranded nucleic acid molecule, and subjecting the cleaved strand(s) to denaturing conditions to remove the portion of the cleaved strand(s) not attached to the solid support, thereby generating a partially or substantially single-stranded template for a nucleic acid sequencing reaction.

摘要翻译： 本发明涉及产生用于核酸测序反应的模板的方法，其包括：提供至少一个双链核酸分子，其中双链核酸分子的两条链在5'末端连接到固体支持物上 切割双链核酸分子的一条或两条链，并使切割的链进行变性条件以除去未连接到固体支持物上的部分切割的链，从而产生部分或基本上 用于核酸测序反应的单链模板。