申请人:LIFE TECHNOLOGIES CORPORATION
公开号:US10329544B2
公开(公告)日:2019-06-25
In some embodiments, the present teachings provide methods for nucleic acid amplification, comprising forming a reaction mixture, and subjecting the reaction mixture to conditions suitable for nucleic acid amplification. In some embodiments, methods for nucleic acid amplification include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, methods for nucleic acid amplification include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the methods for nucleic acid amplification employ an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel. In some embodiments, methods for nucleic acid amplification can be conducted in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components. In some embodiments, methods for nucleic acid amplification comprise a amplifying at least one polynucleotide onto a surface under isothermal amplification conditions, optionally in the presence of a polymer. The polymer can include a sieving agent and/or a diffusion-reducing agent.
在一些实施例中,本教导提供了核酸扩增的方法,包括形成反应混合物,并将反应混合物置于适合核酸扩增的条件下。在一些实施方案中,核酸扩增方法包括将待扩增核酸置于部分变性条件下。在一些实施方案中,核酸扩增的方法包括在不使被扩增核酸完全变性的情况下进行扩增。在某些实施方案中,核酸扩增方法采用催化同源重组的酶和聚合酶。在某些实施方案中,核酸扩增方法可在单个反应容器中进行。在某些实施方案中,核酸扩增方法可在反应混合物的单一连续液相中进行,无需对反应混合物进行分隔或固定反应成分。在某些实施方案中,核酸扩增方法包括在等温扩增条件下,将至少一种多核苷酸扩增到表面上,可选择在有聚合物存在的情况下进行。聚合物可包括筛分剂和/或扩散减少剂。