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使用RNA诱饵对古代DNA进行杂交捕获

Hybridization Capture of Ancient DNA Using RNA Baits.

作者信息

Soares André E R

机构信息

Laboratório Nacional de Computação Científica, Petrópolis, RJ, Brazil.

出版信息

Methods Mol Biol. 2019;1963:121-128. doi: 10.1007/978-1-4939-9176-1_13.

Abstract

The majority of DNA recovered from ancient remains is derived from organisms that colonize the remains post-mortem, such as soil microbes, or from contaminants, such as DNA from living humans. Additionally, some ancient DNA research projects aim to target specific genomic regions, such as mitochondrial genomes or variable single nucleotide polymorphisms (SNPs). To overcome the challenge of targeting specific fragments of DNA from within a complex DNA extract, methods have been developed to enrich ancient DNA extracts for target DNA relative to nontarget DNA. This chapter describes a method for target DNA enrichment that uses hybridization to biotinylated RNA baits to capture and amplify specific ancient DNA fragments from within the pool of extracted fragments.

摘要

从古代遗骸中回收的大部分DNA来自死后在遗骸上定殖的生物体,如土壤微生物,或来自污染物,如活人DNA。此外,一些古代DNA研究项目旨在靶向特定的基因组区域,如线粒体基因组或可变单核苷酸多态性(SNP)。为了克服从复杂的DNA提取物中靶向特定DNA片段的挑战,已经开发出一些方法来相对于非目标DNA富集古代DNA提取物中的目标DNA。本章描述了一种目标DNA富集方法,该方法利用与生物素化RNA诱饵的杂交来从提取的片段池中捕获和扩增特定的古代DNA片段。

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