Preferential access to genetic information from endogenous hominin ancient DNA and accurate quantitative SNP-typing via SPEX
Preferential access to genetic information from endogenous hominin ancient DNA and accurate quantitative SNP-typing via SPEX
复制标题
优先获取内源古人类 DNA 的遗传信息，并通过 SPEX 进行准确的定量 SNP 分型
作者：
Paul Brotherton;J. Sánchez;A. Cooper;P. Endicott
对古代、法医和临床样本中的目标遗传位点的分析通常建立在聚合酶链式反应 (PCR) 生成的序列数据的基础上。然而，许多研究表明，劣质 DNA 模板的 PCR 扩增可能会产生显着水平的序列假象。对于古人类（人类和其他原始人类）样本，绝大多数样本中普遍存在高度 PCR 扩增的人类 DNA 污染物，可能导致产生重组杂交体和其他非真实人工制品。 PCR 生成的序列即使不是不可能，也很难进行验证。相比之下，基于单引物延伸 (SPEX) 的方法可以像现代 DNA 一样准确地对古代 DNA 片段的单核苷酸多态性进行基因分型。单一 SPEX 型检测可以仅扩增目标位点处的双链 DNA 链之一，并产生多倍的覆盖深度，将非真实重组杂交体降低至不可检测的水平。至关重要的是，与现代人类 DNA 污染物相比，SPEX 型方法可以优先从受损和降解的内源性古 DNA 模板中获取遗传信息。 SPEX 型检测的发展为对古代人类样本进行高精度、定量基因分型提供了可能。
The analysis of targeted genetic loci from ancient, forensic and clinical samples is usually built upon polymerase chain reaction (PCR)-generated sequence data. However, many studies have shown that PCR amplification from poor-quality DNA templates can create sequence artefacts at significant levels. With hominin (human and other hominid) samples, the pervasive presence of highly PCR-amplifiable human DNA contaminants in the vast majority of samples can lead to the creation of recombinant hybrids and other non-authentic artefacts. The resulting PCR-generated sequences can then be difficult, if not impossible, to authenticate. In contrast, single primer extension (SPEX)-based approaches can genotype single nucleotide polymorphisms from ancient fragments of DNA as accurately as modern DNA. A single SPEX-type assay can amplify just one of the duplex DNA strands at target loci and generate a multi-fold depth-of-coverage, with non-authentic recombinant hybrids reduced to undetectable levels. Crucially, SPEX-type approaches can preferentially access genetic information from damaged and degraded endogenous ancient DNA templates over modern human DNA contaminants. The development of SPEX-type assays offers the potential for highly accurate, quantitative genotyping from ancient hominin samples.
DOI：
10.1073/pnas.85.23.8998
发表时间：
1988-12-01
影响因子：
11.1
作者：
FROHMAN, MA;DUSH, MK;MARTIN, GR
通讯作者：
MARTIN, GR