公开(公告)号：US20160362737A1

公开(公告)日：2016-12-15

申请号：US15179813

申请日：2016-06-10

申请人： Illumina Cambridge Limited

发明人： John MILTON ,  Xiaolin WU ,  Mark SMITH ,  Joseph BRENNAN ,  Colin BARNES ,  Xiaohai LIU ,  Silke RUEDIGER

IPC分类号： C12Q1/68 ,  C07H19/14 ,  C07H19/10

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.