Horn Susanne
Max Planck Institute for Evolutionary Anthropology, Germany.
Methods Mol Biol. 2012;840:177-88. doi: 10.1007/978-1-61779-516-9_21.
Recent advances in high-throughput DNA sequencing technologies have allowed entire nuclear genomes to be shotgun sequenced from ancient DNA (aDNA) extracts. Nonetheless, targeted analyses of specific genomic loci will remain an important tool for future aDNA studies. DNA capture via hybridization allows the efficient exploitation of current high-throughput sequencing for population genetic analyses using aDNA samples. Specifically, hybridization capture allows larger data sets to be generated for multiple target loci as well as for multiple samples in parallel. "Bait" molecules are used to select target regions from DNA libraries for sequencing. Here we present a brief overview of the currently available hybridization capture protocols using either an in-solution or a solid-phase (immobilized) approach. While it is possible to purchase ready-made kits for this purpose, I present a protocol that allows users to generate their own custom bait to be used for hybridization capture.
高通量DNA测序技术的最新进展使得能够从古代DNA(aDNA)提取物中对整个核基因组进行鸟枪法测序。尽管如此,对特定基因组位点的靶向分析仍将是未来aDNA研究的重要工具。通过杂交进行DNA捕获能够利用当前的高通量测序技术,对aDNA样本进行群体遗传学分析。具体而言,杂交捕获允许针对多个目标位点以及多个样本并行生成更大的数据集。“诱饵”分子用于从DNA文库中选择目标区域进行测序。在此,我们简要概述目前可用的采用溶液内或固相(固定化)方法的杂交捕获方案。虽然可以为此目的购买现成的试剂盒,但我提供了一种方案,允许用户生成自己的定制诱饵用于杂交捕获。