公开(公告)号：US20090269759A1

公开(公告)日：2009-10-29

申请号：US12355487

申请日：2009-01-16

申请人： Steven M. Menchen, JR. ,  Barnett Rosenblum ,  Paul Kenney ,  Shoeb I. Khan ,  Zhaochun Ma ,  Jer-Kang Chen ,  Joe Lam ,  Boli Huang

发明人： Steven M. Menchen, JR. ,  Barnett Rosenblum ,  Paul Kenney ,  Shoeb I. Khan ,  Zhaochun Ma ,  Jer-Kang Chen ,  Joe Lam ,  Boli Huang

IPC分类号： C12Q1/68 ,  C07F9/02

CPC分类号： C07H21/04 ,  C07H19/073 ,  C07H19/173 ,  C12Q1/6874 ,  C12Q2523/107 ,  C12Q2525/186 ,  C12Q2525/197

摘要： Nucleotide analogs that can sustain the enzymatic synthesis of double-stranded nucleic acid from a nucleic template are described. The nucleotide analogs include: (i) a base selected from the group consisting of adenine, guanine, cytosine, thymine, uracil and their analogs; (ii) a label attached to the base or analog of the base via a cleavable linker; (iii) a deoxyribose; and (iv) one or more phosphate groups. The linker and/or the label inhibits template directed polymerase incorporation of a further nucleotide substrate onto an extended primer strand. In addition, cleavage of the linker leaves a residue attached to the base which is not present in the natural nucleotide and which does not inhibit extension of the primer strand. The nucleotide analogs can therefore be used as reversible terminators in sequencing by synthesis methods without blocking the 3′ hydroxyl group. Methods of sequencing DNA using the substrates are also described.

摘要翻译： 描述了可以维持来自核酸模板的双链核酸的酶合成的核苷酸类似物。 核苷酸类似物包括：（i）选自腺嘌呤，鸟嘌呤，胞嘧啶，胸腺嘧啶，尿嘧啶及其类似物的碱基; （ii）通过可切割连接体连接到碱基或类似物的标签; （iii）脱氧核糖; 和（iv）一个或多个磷酸酯基团。 接头和/或标记物将另外的核苷酸底物的模板定向聚合酶掺入到延伸的引物链上。 此外，接头的切割物留下连接到碱基上的残基，其不存在于天然核苷酸中，并且不抑制引物链的延伸。 因此，核苷酸类似物因此可以通过合成方法进行测序而不会阻断3'羟基作为可逆终止子。 还描述了使用底物测序DNA的方法。