Ovchinnikov Igor V, Malek Mathew J, Kjelland Katelyn, Drees Kenneth
Department of Biology, University of North Dakota, Starcher Hall, 10 Cornell Street Stop 9010, Grand Forks, ND, 58202, USA.
Methods Mol Biol. 2016;1420:157-71. doi: 10.1007/978-1-4939-3597-0_13.
Mitochondrial DNA (mtDNA) can help in the identification of biological evidence recovered from crime scenes and human remains. Typically the hypervariable regions are targeted for sequencing; however, more discriminating profiles are obtained if the whole genome is sequenced. Different approaches exist as to how best amplify and sequence whole mtDNA from forensic specimens. Here, we describe a method based on two-round PCR, combining multiplex and simplex PCRs. This method has been used in the analysis of mitochondrial genomes from archival saliva samples applied to FTA® cards after 10 years of transportation and preservation, without special protection. It is expected that this technique can be also used for the analysis of other old biological specimens directly or with modifications related to the level of DNA degradation.
线粒体DNA(mtDNA)有助于识别从犯罪现场和人类遗骸中获取的生物证据。通常,高变区是测序的目标区域;然而,如果对整个基因组进行测序,则可获得更具鉴别力的图谱。关于如何从法医样本中最佳地扩增和测序整个mtDNA,存在不同的方法。在此,我们描述了一种基于两轮PCR的方法,该方法结合了多重PCR和单重PCR。这种方法已用于分析经过10年运输和保存且未进行特殊保护的、应用于FTA®卡的存档唾液样本中的线粒体基因组。预计该技术也可直接用于分析其他陈旧生物样本,或根据DNA降解水平进行修改后使用。