Continuous extension and deblocking in reactions for nucleic acids synthesis and sequencing

    公开(公告)号:US10378051B2

    公开(公告)日:2019-08-13

    申请号:US13627134

    申请日:2012-09-26

    IPC分类号: C40B20/00 C12Q1/68 C12Q1/6874

    摘要: A reaction mixture including (a) a nucleic acid having a primer hybridized to a template, (b) nucleotide analogs, wherein the nucleotide analogs have a blocking moiety; (c) a polymerase that is capable of forming an extended primer by adding the nucleotide analogs to the primer, and (d) a deblocking agent that is capable of removing the blocking moiety from the extended primer. Also provided is a method of synthesizing a polynucleotide including sequentially adding a plurality of the different nucleotides analogs to the nucleic acid via several reaction cycles in the reaction mixture, wherein each reaction cycle includes (i) the polymerase adding a nucleotide analog to the nucleic acid to form a transient nucleic acid species comprising a blocking moiety, and (ii) the deblocking agent modifying the transient nucleic acid species to remove the blocking moiety.

    CONTINUOUS EXTENSION AND DEBLOCKING IN REACTIONS FOR NUCLLEIC ACIDS SYNTHESIS AND SEQUENCING
    8.
    发明申请
    CONTINUOUS EXTENSION AND DEBLOCKING IN REACTIONS FOR NUCLLEIC ACIDS SYNTHESIS AND SEQUENCING 审中-公开
    连续延伸和解除核酸合成和测序的反应

    公开(公告)号:US20130085073A1

    公开(公告)日:2013-04-04

    申请号:US13627134

    申请日:2012-09-26

    摘要: A reaction mixture including (a) a nucleic acid having a primer hybridized to a template, (b) nucleotide analogs, wherein the nucleotide analogs have a blocking moiety; (c) a polymerase that is capable of forming an extended primer by adding the nucleotide analogs to the primer, and (d) a deblocking agent that is capable of removing the blocking moiety from the extended primer. Also provided is a method of synthesizing a polynucleotide including sequentially adding a plurality of the different nucleotides analogs to the nucleic acid via several reaction cycles in the reaction mixture, wherein each reaction cycle includes (i) the polymerase adding a nucleotide analog to the nucleic acid to form a transient nucleic acid species comprising a blocking moiety, and (ii) the deblocking agent modifying the transient nucleic acid species to remove the blocking moiety.

    摘要翻译: 一种反应混合物,其包含(a)具有与模板杂交的引物的核酸,(b)核苷酸类似物,其中所述核苷酸类似物具有阻断部分; (c)通过向引物中加入核苷酸类似物,能够形成延伸引物的聚合酶,和(d)能够从延伸引物除去阻断部分的解封剂。 还提供了合成多核苷酸的方法,所述多核苷酸包括在反应混合物中通过几个反应循环顺序加入多个不同的核苷酸类似物,其中每个反应循环包括(i)将核苷酸类似物加入到核酸中 以形成包含阻断部分的瞬时核酸物质,和(ii)修饰瞬态核酸物质以去除阻断部分的解封端剂。