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案例研究:通过杂交捕获富集古代线粒体DNA

Case study: enrichment of ancient mitochondrial DNA by hybridization capture.

作者信息

Horn Susanne

机构信息

Max Planck Institute for Evolutionary Anthropology, Germany.

出版信息

Methods Mol Biol. 2012;840:189-95. doi: 10.1007/978-1-61779-516-9_22.

Abstract

In ancient DNA studies focusing on estimating population histories, genetic markers are sequenced from a large number of samples belonging to the same species. Targeting loci of interest using traditional PCR can be time-consuming, in particular when samples are not well preserved and multiple overlapping fragments are required. Here, I describe the process of generating DNA libraries from ancient DNA (aDNA) extracts for high-throughput sequencing. I use a serial in-solution DNA hybridization approach with subsequent bead capture to enrich libraries for the target locus, in this case the mitochondrial control region of ancient beavers (Castor fiber). The resulting sequencing reads are run through quality control filters to obtain reliable consensus sequences. Using these sequences, I construct a phylogenetic tree, which agrees with previously published data regarding phylogeographic relationships among beavers.

摘要

在专注于估计种群历史的古DNA研究中,从属于同一物种的大量样本中对遗传标记进行测序。使用传统PCR靶向感兴趣的位点可能很耗时,特别是当样本保存不佳且需要多个重叠片段时。在这里,我描述了从古代DNA(aDNA)提取物中生成DNA文库以进行高通量测序的过程。我使用连续的溶液内DNA杂交方法,随后进行磁珠捕获,以富集文库中的目标位点,在这种情况下是古代河狸(Castor fiber)的线粒体控制区。将得到的测序读数通过质量控制过滤器以获得可靠的共有序列。使用这些序列,我构建了一个系统发育树,它与先前发表的关于河狸系统地理关系的数据一致。

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