申请人:LIFE TECHNOLOGIES CORPORATION
公开号:US11001815B2
公开(公告)日:2021-05-11
The present disclosure provides methods, compositions, kits and systems for nucleic acid amplification. In some embodiments, nucleic acid amplification methods include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, nucleic acid amplification methods include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the nucleic acid amplification method employs 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 and/or 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 amplifying at least one polynucleotide onto a surface under isothermal amplification conditions, optionally in the presence of a polymer which can include a sieving agent and/or a diffusion-reducing agent.
本公开提供了用于核酸扩增的方法、组合物、试剂盒和系统。在某些实施方案中,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。在一些实施方案中,核酸扩增方法采用催化同源重组的酶和聚合酶。在一些实施方案中,核酸扩增方法可在单个反应容器和/或反应混合物的单个连续液相中进行,无需对反应混合物进行分隔或固定反应组分。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。