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：US20090062145A1

公开(公告)日：2009-03-05

申请号：US12220682

申请日：2008-07-24

申请人： Xiaohai Liu ,  John Milton ,  Silke Ruediger

发明人： Xiaohai Liu ,  John Milton ,  Silke Ruediger

IPC分类号： C40B40/08

CPC分类号： C07H19/04 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C07H21/04 ,  C12Q1/6869 ,  C12Q1/6874 ,  Y10T436/143333 ,  C12Q2535/113 ,  C12Q2525/186

摘要： The invention provides a nucleotide or nucleoside having a base attached to a detectable label via a cleavable linker, characterised in that the cleavable linker contains a moiety selected from the group comprising: Formula (I) (wherein X is selected from the group comprising O, S, NH and NQ wherein Q is a C1-10 substituted or unsubstituted alkyl group, Y is selected from the group comprising O, S, NH and N(allyl). T is hydrogen or a C1-10 substituted or unsubstituted alkyl group and * indicates where the moiety is connected to the remainder of the nucleotide or nucleoside).

摘要翻译： 本发明提供了具有通过可切割接头连接到可检测标记的碱基的核苷酸或核苷，其特征在于所述可切割连接体含有选自以下的部分：式（I）（其中X选自O， S，NH和NQ，其中Q是C1-10取代或未取代的烷基，Y选自O，S，NH和N（烯丙基），T是氢或C1-10取代或未取代的烷基， *表示部分连接到核苷酸或核苷的剩余部分）。

公开(公告)号：US20060188901A1

公开(公告)日：2006-08-24

申请号：US11301578

申请日：2005-12-13

申请人： Colin Barnes ,  Shankar Balasubramanian ,  Xiaohai Liu ,  Harold Swerdlow ,  John Milton

发明人： Colin Barnes ,  Shankar Balasubramanian ,  Xiaohai Liu ,  Harold Swerdlow ,  John Milton

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

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

摘要： Nucleosides and nucleotides are disclosed that are linked to detectable labels via a cleavable linker group.

公开(公告)号：US20140349891A1

公开(公告)日：2014-11-27

申请号：US14166670

申请日：2014-01-28

申请人： Xiaohai Liu ,  John Milton ,  Geoffrey Paul Smith ,  Colin Lloyd 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 Lloyd 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： C12Q1/68

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，

公开(公告)号：US08212015B2

公开(公告)日：2012-07-03

申请号：US12899423

申请日：2010-10-06

申请人： John Milton ,  Xiaohai Liu

发明人： John Milton ,  Xiaohai Liu

IPC分类号： C07H21/04 ,  C07H19/04 ,  C12Q1/68 ,  C12P19/34 ,  B01L3/00

CPC分类号： C12Q1/686 ,  C12Q2525/113

摘要： The invention is directed to modified guanine-containing nucleosides and nucleotides and uses thereof. More specifically, the invention relates to modified fluorescently labelled guanine-containing nucleosides and nucleotides which exhibit enhanced fluorophore intensity by virtue of reduced quenching effects.

摘要翻译： 本发明涉及修饰的含鸟嘌呤核苷及其用途。 更具体地，本发明涉及经修饰的荧光标记的含有鸟嘌呤的核苷和核苷酸，其通过降低猝灭效应表现出增强的荧光团强度。

公开(公告)号：US20100311178A1

公开(公告)日：2010-12-09

申请号：US12803163

申请日：2010-06-21

申请人： Xiaohai Liu ,  John Milton ,  Silke Ruediger

发明人： Xiaohai Liu ,  John Milton ,  Silke Ruediger

IPC分类号： G01N33/50 ,  C07H19/10 ,  C07H21/04

CPC分类号： C07H19/04 ,  C07H19/06 ,  C07H19/10 ,  C07H19/14 ,  C07H19/16 ,  C07H19/20 ,  C07H21/00 ,  C07H21/04 ,  C12Q1/6869 ,  C12Q1/6874 ,  Y10T436/143333 ,  C12Q2535/113 ,  C12Q2525/186

摘要： The invention provides a nucleotide or nucleoside having a base attached to a detectable label via a cleavable linker, characterised in that the cleavable linker contains a moiety selected from the group comprising: Formula (I) (wherein X is selected from the group comprising O, S, NH and NQ wherein Q is a C1-10 substituted or unsubstituted alkyl group, Y is selected from the group comprising O, S, NH and N(allyl). T is hydrogen or a C1-10 substituted or unsubstituted alkyl group and * indicates where the moiety is connected to the remainder of the nucleotide or nucleoside).

摘要翻译： 本发明提供了具有通过可切割接头连接到可检测标记的碱基的核苷酸或核苷，其特征在于所述可切割连接体含有选自以下的部分：式（I）（其中X选自O， S，NH和NQ，其中Q是C1-10取代或未取代的烷基，Y选自O，S，NH和N（烯丙基），T是氢或C1-10取代或未取代的烷基， *表示部分连接到核苷酸或核苷的剩余部分）。

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：US07816503B2

公开(公告)日：2010-10-19

申请号：US12586385

申请日：2009-09-21

申请人： John Milton ,  Xiaohai Liu

发明人： John Milton ,  Xiaohai Liu

IPC分类号： C07H21/00 ,  C07H21/02 ,  C07H21/04 ,  C12Q1/68 ,  C12P19/34

CPC分类号： C12Q1/686 ,  C12Q2525/113

摘要： The invention is directed to modified guanine-containing nucleosides and nucleotides and uses thereof. More specifically, the invention relates to modified fluorescently labelled guanine-containing nucleosides and nucleotides which exhibit enhanced fluorophore intensity by virtue of reduced quenching effects.

摘要翻译： 本发明涉及修饰的含鸟嘌呤核苷及其用途。 更具体地，本发明涉及经修饰的荧光标记的含有鸟嘌呤的核苷和核苷酸，其通过降低猝灭效应表现出增强的荧光团强度。

公开(公告)号：US20100068722A1

公开(公告)日：2010-03-18

申请号：US12586385

申请日：2009-09-21

申请人： John Milton ,  Xiaohai Liu

发明人： John Milton ,  Xiaohai Liu

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12Q1/686 ,  C12Q2525/113

摘要： The invention is directed to modified guanine-containing nucleosides and nucleotides and uses thereof. More specifically, the invention relates to modified fluorescently labelled guanine-containing nucleosides and nucleotides which exhibit enhanced fluorophore intensity by virtue of reduced quenching effects.

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.