Simple and highly effective DNA extraction methods from old skeletal remains using silica columns
Simple and highly effective DNA extraction methods from old skeletal remains using silica columns
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DOI：
10.1016/j.fsigen.2009.10.014
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发表时间：
2010-10-01
影响因子：
3.1
通讯作者：
Shin, Kyoung-Jin
中科院分区：
文献类型：
--
作者：
Lee, Hwan Young;Park, Myung Jin;Shin, Kyoung-Jin
由于提取的DNA的降解和非常低的产量以及PCR抑制剂的存在，从旧骨骼遗骸中恢复DNA数据通常是困难的。在此，我们比较了几种基于二氧化硅的DNA提取方法从人工降解的DNA，DNA与PCR抑制剂和DNA从旧的骨骼遗骸使用定量实时PCR。我们提出了一种改进的大规模基于二氧化硅的提取与完全脱矿相结合，使最大的DNA回收和有效消除PCR抑制剂。这是用高浓度EDTA溶液进行骨粉脱矿，然后用QIAamp离心柱和来自QIAquick PCR纯化试剂盒的缓冲液进行的。我们已经成功地使用这种改进的技术进行STR分析的55岁的骨骼遗骸。本文的研究结果将有助于解决法医学处理遗骸案件。(C)2010爱思唯尔爱尔兰有限公司版权所有。
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 (R) spin columns and buffers from the QIAquick (R) 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. (C) 2010 Elsevier Ireland Ltd. All rights reserved.