Centre for Virology, Burnet Institute, 85 Commercial Road, Melbourne, Victoria, Australia.
Gene. 2010 Dec 1;469(1-2):45-51. doi: 10.1016/j.gene.2010.08.009. Epub 2010 Sep 15.
Measurements of population diversity are fundamental to the reconstruction of the evolutionary and epidemiological history of organisms. Commonly used protocols to measure population diversity using the polymerase chain reaction (PCR) are prone to the introduction of artificial chimeras. These are often difficult to detect and can confound the correct interpretation of results due to the false generation of recombinants when the underlying DNA sample contains multiple distinct templates. This study presents a standardised procedure to suppress the formation of artificial chimeras during PCR amplification. The solution is based on the accurate determination of the efficiency and end point of the log-linear phase of a PCR. This procedure will facilitate the generation of data sets that more accurately reflect the underlying population diversity rather than artifacts introduced by the process itself.
种群多样性的衡量是重建生物进化和流行病学历史的基础。聚合酶链反应（PCR）是常用的测量种群多样性的方法，但该方法容易产生人为嵌合体。由于当基础 DNA 样本包含多个不同模板时，错误地产生了重组体，因此这些嵌合体通常很难被检测到，并且会使结果的正确解释变得复杂。本研究提出了一种在 PCR 扩增过程中抑制人为嵌合体形成的标准化程序。该解决方案基于对 PCR 对数线性阶段的效率和终点的准确确定。该程序将有助于生成更准确地反映基础种群多样性的数据，而不是由该过程本身引入的假象。
