Piglionica Marilidia, De Donno Antonio, Baldassarra Stefania Lonero, Santoro Valeria, Scorca Antonella, Introna Francesco, Dell'Erba Alessandro
Section of Legal Medicine, Bari, Italy.
Am J Forensic Med Pathol. 2012 Dec;33(4):322-7. doi: 10.1097/PAF.0b013e3182519479.
To resolve cases involving unidentified cadavers, the study of polymorphic DNA markers of old bones is an invaluable but often challenging tool used in forensic genetics. Some of the difficulties encountered involve the limited quantity of endogenous DNA, its subsequent degradation (a result of elapsed time, environmental conditions, and the microorganisms that develop during the postmortem phase), and the coextraction of substances that inhibit amplification reactions. For these reasons, it is necessary to direct research toward the development of new extraction techniques with the goal of obtaining adequate quantities of high-quality DNA.The aim of this study was to improve the collection of extracted DNA compared with the amount of DNA obtained with the NucleoSpin DNA Trace Kit (Macherey Nagel) protocol for the extraction of genomic DNA from human bones. A modified version of the standard protocol is presented.The modified method for the extraction of genomic DNA, followed by amplification reaction, allowed for identification of 4 cadavers and typification of 1 cadaver. The study carried out involved unidentified cadavers, or their remains, discovered after a long period from time of death.
为了解决涉及身份不明尸体的案件,对陈旧骨骼的多态性DNA标记进行研究是法医遗传学中一种非常宝贵但通常具有挑战性的工具。遇到的一些困难包括内源性DNA数量有限、随后的降解(这是死亡时间、环境条件以及死后阶段滋生的微生物导致的结果),以及抑制扩增反应的物质的共提取。出于这些原因,有必要将研究方向转向开发新的提取技术,目标是获得足够数量的高质量DNA。本研究的目的是与使用NucleoSpin DNA Trace试剂盒(Macherey Nagel)从人骨中提取基因组DNA的方案相比,改进提取DNA的收集量。本文介绍了标准方案的修改版本。经修改的基因组DNA提取方法,随后进行扩增反应,能够鉴定4具尸体,并对1具尸体进行分型。所开展的研究涉及身份不明的尸体或其遗骸,这些尸体或遗骸是在死后很长一段时间才被发现的。