Department of Forensic Medicine and Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, 250 Seongsanno, Seodaemun-Gu, Seoul 120-752, Republic of Korea.
Forensic Sci Int Genet. 2010 Oct;4(5):275-80. doi: 10.1016/j.fsigen.2009.10.014. Epub 2010 Jan 19.
The recovery of DNA data from old skeletal remains is often difficult due to degraded and very low yield of extracted DNA and the presence of PCR inhibitors. Herein, we compared several silica-based DNA extraction methods from artificially degraded DNA, DNA with PCR inhibitors and DNA from old skeletal remains using quantitative real-time PCR. We present a modified large-scale silica-based extraction combined with complete demineralization, that enables maximum DNA recovery and efficient elimination of PCR inhibitors. This is performed with high concentration of EDTA solution for demineralization of bone powder followed by QIAamp spin columns and buffers from the QIAquick PCR purification kit. We have successfully used this modified technique to perform STR analysis for 55-year-old skeletal remains. The results of this study will contribute to solve the forensic cases dealing with skeletal remains.
从陈旧的骨骼遗骸中恢复 DNA 数据通常很困难,因为提取的 DNA 降解严重且产量非常低,并且存在 PCR 抑制剂。在此,我们使用定量实时 PCR 比较了几种基于硅的从人工降解的 DNA、带有 PCR 抑制剂的 DNA 和陈旧骨骼遗骸中提取 DNA 的方法。我们提出了一种改良的大型基于硅的提取方法,该方法结合了完全脱矿化,可实现最大程度的 DNA 回收并有效消除 PCR 抑制剂。这是通过高浓度 EDTA 溶液对骨粉进行脱矿质处理,然后使用 QIAamp 离心柱和 QIAquick PCR 纯化试剂盒中的缓冲液来完成的。我们已经成功地使用这种改良技术对 55 岁的骨骼遗骸进行 STR 分析。这项研究的结果将有助于解决涉及骨骼遗骸的法医案例。