Abstract：
Identification of human remains using genetic methods is an important task of forensic science. DNA markers are proving essential in the identification of unknown human remains. However, environmental factors can lead to poor preservation of DNA, including in bone material. The aim of this study was therefore to compare two methods of DNA isolation from bone material: the traditional organic method and the new protocol using the EZ2 Connect instrument. The study involved three types of bone material, namely molars/premolars, petrous parts of the temporal bone and femurs, all with an estimated PMI of 70-80 years. Importantly, the biological material was obtained from three different environments, categorized as preserving, neutral and degrading, based on basic physico-chemical tests and the potential impact on the bone. The results obtained show that the DNA was best preserved in the petrous bone, followed by the teeth, and the femur. DNA extraction using the EZ2 Connect instrument with a new protocol gave slightly better results for the petrous bone, comparable results for the teeth and worse results for the femur compared to the organic method. Several protocol modifications were tested and optimal conditions for DNA isolation were proposed for the EZ2 protocol. Furthermore, the use of an automated method facilitated the effective accumulation of isolates and increased the chances of successful identification of unknown human remains.
摘要：
使用遗传方法鉴定人类遗骸是法医学的重要任务。DNA标记在鉴定未知的人类遗骸中被证明是必不可少的。然而,环境因素会导致DNA保存不良，包括骨骼材料。因此，这项研究的目的是比较两种从骨材料中分离DNA的方法：传统的有机方法和使用EZ2Connect仪器的新方案。这项研究涉及三种类型的骨材料，即磨牙/前磨牙，颞骨和股骨的岩质部分，估计PMI为70-80年。重要的是,生物材料来自三种不同的环境，归类为保存，中立和有辱人格的，基于基本的物理化学测试和对骨骼的潜在影响。获得的结果表明，DNA最好保存在岩骨中，接着是牙齿，还有股骨.使用具有新方案的EZ2Connect仪器提取的DNA对岩骨的结果略好，与有机方法相比，牙齿的结果相当，股骨的结果较差。测试了几种方案修改，并为EZ2方案提出了DNA分离的最佳条件。此外，自动化方法的使用促进了分离株的有效积累，并增加了成功识别未知人类遗骸的机会。
