申请人:Mir Kalim U.
公开号:US11066701B2
公开(公告)日:2021-07-20
A method for template-directed sequencing-by-synthesis of an array of target polynucleotide can include:
(a) providing an array of target polynucleotides in a fluidic vessel;
(b) contacting the array of polynucleotides with a solution comprising (i) polymerization complex and (ii) reversibly terminating and differently labeled A,C,G, and T/U nucleotides;
(c) incorporating one of the differently labeled nucleotides, using the polymerization complex, into a chain complementary to at least one of the array of polynucleotides;
(d) binding imaging tags to the differently labeled nucleotides of step (c);
(e) imaging and storing the identity and position of the imaging tags of step (d);
(f) reversing termination (b)-(e);
(g) repeating steps (b)-(e) and assembling a sequence for each of the array of target polynucleotides from the stored identity and position of the imaging tags, optionally as a homogeneous or one pot reaction. Additional methods of sequencing target polynucleotides are described herein.
通过合成对目标多核苷酸阵列进行模板定向测序的方法可包括
(a) 在流体容器中提供目标多核苷酸阵列;
(b) 将多核苷酸阵列与溶液接触,溶液包括:(i) 聚合复合物;(ii) 可逆终止和不同标记的 A、C、G 和 T/U 核苷酸;
(c) 使用聚合复合物将其中一种不同标记的核苷酸结合到与多核苷酸阵列中至少一种核苷酸互补的链中;
(d) 将成像标签与步骤(c)中不同标记的核苷酸结合;
(e) 对步骤(d)的成像标签的身份和位置进行成像和存储;
(f) 反转步骤(b)-(e)
(g) 重复步骤(b)-(e),并根据存储的成像标记的身份和位置,为每个目标多核苷酸阵列组装序列,可选择作为均相反应或一锅反应。本文还描述了对目标多核苷酸进行测序的其他方法。