Forst Jannine, Brown Terence A
Manchester Institute of Biotechnology, School of Earth and Environmental Sciences, University of Manchester, Manchester, United Kingdom.
PLoS One. 2016 Sep 21;11(9):e0163031. doi: 10.1371/journal.pone.0163031. eCollection 2016.
We assessed the ability of whole genome amplification (WGA) to improve the efficiency of downstream polymerase chain reactions (PCRs) directed at ancient DNA (aDNA) of members of the Mycobacterium tuberculosis complex (MTBC). Using extracts from a variety of bones and a tooth from human skeletons with or without lesions indicative of tuberculosis, from multiple time periods, we obtained inconsistent results. We conclude that WGA does not provide any advantage in studies of MTBC aDNA. The sporadic nature of our results are probably due to the fact that WGA is itself a PCR-based procedure which, although designed to deal with fragmented DNA, might be inefficient with the low concentration of templates in an aDNA extract. As such, WGA is subject to similar, if not the same, restrictions as PCR when applied to aDNA.
我们评估了全基因组扩增(WGA)提高针对结核分枝杆菌复合群(MTBC)成员的古代DNA(aDNA)进行下游聚合酶链反应(PCR)效率的能力。我们使用了来自多个时期、有或没有结核病病变迹象的人类骨骼的各种骨骼和一颗牙齿的提取物,结果并不一致。我们得出结论,WGA在MTBC aDNA研究中没有提供任何优势。我们结果的零散性质可能是由于WGA本身是一种基于PCR的程序,尽管它旨在处理片段化的DNA,但对于aDNA提取物中低浓度的模板可能效率低下。因此,当应用于aDNA时,WGA受到与PCR类似(如果不是相同)的限制。