Abstract
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.
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Acknowledgments
The study was supported by the North Dakota EPSCoR and UND faculty start-up awards to I.V.O. The research protocol was approved by the University of North Dakota’s Institutional Review Board (Protocol number IRB-201307-009) and the University of North Dakota’s Institutional Biosafety Committee (Registration number IBC-201102-001).
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Ovchinnikov, I.V., Malek, M.J., Kjelland, K., Drees, K. (2016). Whole Human Mitochondrial DNA Sequencing. In: Goodwin, W. (eds) Forensic DNA Typing Protocols. Methods in Molecular Biology, vol 1420. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3597-0_13
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DOI: https://doi.org/10.1007/978-1-4939-3597-0_13
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