Letts Brandon, Shapiro Beth
Department of Biology, The Pennsylvania State University, 320 Mueller Laboratory, University Park, PA 16802, USA.
Methods Mol Biol. 2012;840:87-92. doi: 10.1007/978-1-61779-516-9_12.
Warm, humid regions are not ideal for long-term DNA preservation. Consequently, little ancient DNA research has been carried out involving taxa that lived in, for example, tropical and subtropical regions. Those studies that have isolated ancient DNA from warm environments have mostly been limited to the most recent several thousand years. Here, we discuss an ancient DNA experiment in which we attempt to amplify mitochondrial DNA from remains of armadillo, glyptodont, and pampathere from sites in Florida, USA, all believed to be around 10,000-12,000 years old. We were successful in recovering DNA from only one of these samples. However, based on the amount and distribution of DNA damage, the ancient DNA recovered was well-preserved despite the age and preservation environment. In this case study chapter, we discuss the experimental procedure we used to characterize the DNA from the Floridian samples, focusing on challenges of working with ancient specimens from warm environments and steps taken to confirm the authenticity of the recovered sequence.
温暖潮湿的地区并非长期保存DNA的理想环境。因此,涉及生活在热带和亚热带等地区的生物分类群的古代DNA研究开展得很少。那些从温暖环境中分离出古代DNA的研究大多局限于最近的几千年。在此,我们讨论一项古代DNA实验,在该实验中,我们试图从美国佛罗里达州一些遗址中犰狳、雕齿兽和潘帕兽的遗骸中扩增线粒体DNA,这些遗址的年代均被认为在10000至12000年左右。我们仅从其中一个样本中成功提取到了DNA。然而,基于DNA损伤的数量和分布情况,尽管样本年代久远且保存环境不佳,但所提取的古代DNA保存状况良好。在本案例研究章节中,我们将讨论用于鉴定佛罗里达样本DNA的实验过程,重点关注处理来自温暖环境的古代样本所面临的挑战以及为确认所提取序列的真实性而采取的步骤。