Armed Forces Medical Examiner System's Armed Forces DNA Identification Laboratory (AFMES-AFDIL), Dover Air Force Base, Dover, DE, USA; SNA International LLC, Alexandria, VA, USA.
Armed Forces Medical Examiner System's Armed Forces DNA Identification Laboratory (AFMES-AFDIL), Dover Air Force Base, Dover, DE, USA; SNA International LLC, Alexandria, VA, USA; Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
Forensic Sci Int Genet. 2021 Jul;53:102496. doi: 10.1016/j.fsigen.2021.102496. Epub 2021 Mar 13.
In the past decade, hybridization capture has gained attention within the forensic field for its possible use in human identification. One of the primary benefits to capture enrichment is its applicability to degraded DNA fragments that, due to their reduced size, are not amenable to traditional PCR enrichment techniques. Hybridization capture is typically introduced after genomic library preparation of extracted DNA templates for the subsequent enrichment of mitochondrial DNA or single nucleotide polymorphisms within the nuclear genome. The enriched molecules are then subjected to massively parallel sequencing (MPS) for sensitive and high-throughput DNA sequence generation. Bioinformatic analysis of capture product removes PCR duplicates that were introduced during the laboratory workflow in order to characterize the original DNA template molecules. In the case of aged and degraded skeletal remains, the fraction of endogenous human DNA may be very low; therefore low-coverage sequence analysis may be required. This review contains an overview of current capture methodologies and the primary literature on hybridization capture as evaluated for forensic applications.
在过去的十年中,杂交捕获技术在法医领域引起了关注,因为它可能用于人类识别。捕获富集的一个主要优点是,它适用于因尺寸减小而不适合传统 PCR 富集技术的降解 DNA 片段。杂交捕获通常在提取 DNA 模板的基因组文库制备后进行,以便随后富集线粒体 DNA 或核基因组中的单核苷酸多态性。富集的分子随后进行大规模平行测序(MPS),以进行敏感和高通量的 DNA 序列生成。捕获产物的生物信息学分析去除了在实验室工作流程中引入的 PCR 重复,以便对原始 DNA 模板分子进行特征描述。在陈旧和降解的骨骼遗骸的情况下,内源性人类 DNA 的比例可能非常低;因此,可能需要进行低覆盖率序列分析。本综述包含对当前捕获方法的概述以及对法医应用进行评估的杂交捕获主要文献。