School of Marine Science and Technology, Harbin Institute of Technology, Weihai 264209, China.
Institute of Biological Sciences and Biotechnology, College of Chemistry, Chemical Engineering and Biotechnology, Donghua University, Shanghai 201620, China
Acta Biochim Biophys Sin (Shanghai). 2014 Jun;46(6):502-11. doi: 10.1093/abbs/gmu026. Epub 2014 Apr 24.
Quantum dots (QDs) are of great interest due to their unique chemical and physical properties. Recently, a hot start (HS) polymerase chain reaction (PCR) amplification performance based on QDs with a high-fidelity Pfu DNA polymerase has been reported. However, whether QDs can trigger HS effects with other high-fidelity or conventional DNA polymerases is yet to be understood. In the present study, we studied the QD-triggered HS effects with four high-fidelity and three conventional DNA polymerases, and the HS effect comparisons among them were also made. It was found that QDs could trigger a distinct HS PCR amplification performance with all the four tested high-fidelity DNA polymerases, and specific target DNA could be well amplified even if the PCR mixture was pre-incubated for 2 h at 50°C. On the contrary, the HS effects were not prominent with all the three conventional Taq DNA polymerases. Specifically, the fidelity of Pfu is not sacrificed in the presence of QDs, even after a 1 h pre-incubation at 50°C before PCR. Furthermore, the electrophoresis results preliminarily demonstrated that QDs prefer to adsorb high-fidelity polymerases rather than conventional ones, which might result in the QD-triggered HS effects on PCR performance by using high-fidelity DNA polymerases.
量子点 (QDs) 因其独特的化学和物理性质而备受关注。最近，据报道，一种基于具有高保真 Pfu DNA 聚合酶的 QD 的热启动 (HS) 聚合酶链反应 (PCR) 扩增性能。然而，QDs 是否可以与其他高保真或常规 DNA 聚合酶触发 HS 效应尚不清楚。在本研究中，我们研究了四种高保真和三种常规 DNA 聚合酶触发的 QD 引发的 HS 效应，并对它们之间的 HS 效应进行了比较。结果发现，QDs 可以触发所有四种测试的高保真 DNA 聚合酶的明显 HS PCR 扩增性能，即使在 50°C 下预孵育 2 小时，也可以很好地扩增特定的靶 DNA。相反，所有三种常规 Taq DNA 聚合酶都没有明显的 HS 效应。具体来说，即使在 PCR 之前在 50°C 下预孵育 1 小时，Pfu 的保真度也不会在 QD 的存在下受到牺牲。此外，电泳结果初步表明，QDs 更喜欢吸附高保真聚合酶而不是常规聚合酶，这可能导致使用高保真 DNA 聚合酶对 PCR 性能产生 QD 触发的 HS 效应。
