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使用自制诱饵对古代DNA进行全基因组捕获

Whole-Genome Capture of Ancient DNA Using Homemade Baits.

作者信息

González Fortes Gloria, Paijmans Johanna L A

机构信息

Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.

Institute for Biochemistry and Biology, University of Potsdam, Potsdam, Germany.

出版信息

Methods Mol Biol. 2019;1963:93-105. doi: 10.1007/978-1-4939-9176-1_11.

Abstract

For many archaeological and paleontological samples, the relative content of endogenous compared to contaminant DNA is low. In such cases, enriching sequencing libraries for endogenous DNA, prior to sequencing can make the final research project more cost-effective. Here, we present an in-solution enrichment protocol based on homemade baits that can be applied to recover complete nuclear genomes from ancient remains. The approach is based on the preparation of DNA baits by biotinylated adapter ligation. The procedure has been developed for use with human remains but can be adapted to other species or target regions by choosing the appropriate template DNA from which to build the capture baits. By using homemade rather than commercially acquired baits, this protocol may offer increased flexibility and cost efficiency.

摘要

对于许多考古和古生物学样本而言,与污染DNA相比,内源性DNA的相对含量较低。在这种情况下,在测序之前对内源性DNA的测序文库进行富集可以使最终的研究项目更具成本效益。在此,我们提出了一种基于自制诱饵的溶液内富集方案,该方案可用于从古遗骸中恢复完整的核基因组。该方法基于通过生物素化接头连接制备DNA诱饵。此程序已开发用于人类遗骸,但通过选择构建捕获诱饵的合适模板DNA,可适用于其他物种或目标区域。通过使用自制而非商业购买的诱饵,该方案可能会提供更高的灵活性和成本效益。

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