In recent years, methods of isothermal amplification of nucleic acids using polymerases with a strand-displacement activity have become widespread for identification of specific nucleotide sequences. Bst exo– polymerase is the most popular of these polymerases, although it is inclined to nonspecific amplification (the so-called multimerization), which leads to the accumulation of by-products constructed of tandem nucleotide repeats. In this study, we evaluated the efficiency of multimerization depending on the reaction conditions and proposed some methods for its elimination. The highest efficiency of multimerization was found in the case of Bst 2.0 polymerase in Isothermal buffer, whereas the Bst-like Gss polymerase provided the formation of multimerization products only in Isothermal buffer and at the latest stages of the reaction. The optimal method for elimination of multimerization was the use of Gss polymerase and Thermopol buffer, or Bst LF polymerase and Isothermal II buffer, or Bst 3.0 polymerase and Thermopol buffer, or Bst 3.0 polymerase in Isothermal buffer and Mn2+ ions as a cofactor. In these cases specific isothermal amplification of the target DNA may take place and provide accurate and reliable results.

中文翻译：

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消除 Bst Exo-DNA 聚合酶存在下等温扩增引起的 DNA 多聚化

近年来，使用具有链置换活性的聚合酶等温扩增核酸的方法已广泛用于鉴定特定核苷酸序列。Bst 外切聚合酶是这些聚合酶中最受欢迎的，尽管它倾向于非特异性扩增（所谓的多聚化），这会导致由串联核苷酸重复构成的副产物积累。在这项研究中，我们根据反应条件评估了多聚化的效率，并提出了一些消除多聚化的方法。Bst 2.0 聚合酶在等温缓冲液中的多聚化效率最高，而 Bst 样 Gss 聚合酶仅在等温缓冲液中和反应的最新阶段提供多聚化产物的形成。消除多聚化的最佳方法是使用 Gss 聚合酶和 Thermopol 缓冲液，或 Bst LF 聚合酶和等温 II 缓冲液，或 Bst 3.0 聚合酶和 Thermopol 缓冲液，或等温缓冲液中的 Bst 3.0 聚合酶和 Mn2+ 离子作为辅助因子。在这些情况下，可能会发生目标 DNA 的特定等温扩增并提供准确可靠的结果。