Abstract
The emergence of Massively Parallel Sequencing technologies enabled the analysis of full mitochondrial (mt)DNA sequences from forensically relevant samples that have, so far, only been typed in the control region or its hypervariable segments. In this study, we evaluated the performance of a commercially available multiplex-PCR-based assay, the Precision ID mtDNA Whole Genome Panel (Thermo Fisher Scientific), for the amplification and sequencing of the entire mitochondrial genome (mitogenome) from even degraded forensic specimens. For this purpose, more than 500 samples from 24 different populations were selected to cover the vast majority of established superhaplogroups. These are known to harbor different signature sequence motifs corresponding to their phylogenetic background that could have an effect on primer binding and, thus, could limit a broad application of this molecular genetic tool. The selected samples derived from various forensically relevant tissue sources and were DNA extracted using different methods. We evaluated sequence concordance and heteroplasmy detection and compared the findings to conventional Sanger sequencing as well as an orthogonal MPS platform. We discuss advantages and limitations of this approach with respect to forensic genetic workflow and analytical requirements.
| Original language | English |
|---|---|
| Pages (from-to) | 244-251 |
| Number of pages | 8 |
| Journal | Forensic Science International: Genetics |
| Volume | 42 |
| DOIs | |
| State | Published - Sep 2019 |
Bibliographical note
Funding Information:This work was supported in part by the European Union grant agreement number 779485-STEFA - ISFP-2016-AG-IBA-ENFSI , CAPES Pro-Forense grant number 23038.006844/2014-46 , the "Theodor-Körner-Fonds zur Förderung von Wissenschaft und Kunst" grant number 3481 , the National Institute of Justice , Office of Justice Programs , U.S. Department of Justice grant number 2016-DN-BX-K001 , and the Tiroler Wissenschaftsfonds (TWF) ( UNI-404/1998 ). We would like to thank Daniel Corach (University of Buenos Aires), Alessandro Achilli (University of Pavia, Italy) and Hovirag Lancioni, (University of Perugia) for providing samples. We would like to thank Tatiana Nogueira and Abdul A. Al-Deib for their technical assistance in obtaining the Brazilian Native American and Libyan samples, respectively, used in this study. We also would like to thank Maiko Takahashi for her technical assistance in this study. The opinions, findings, and conclusions or recommendations expressed in this publication are those of the authors and do not necessarily reflect those of the U.S. Department of Justice.
Funding Information:
This work was supported in part by the European Union grant agreement number 779485-STEFA - ISFP-2016-AG-IBA-ENFSI, CAPES Pro-Forense grant number 23038.006844/2014-46, the ?Theodor-K?rner-Fonds zur F?rderung von Wissenschaft und Kunst? grant number 3481, the National Institute of Justice, Office of Justice Programs, U.S. Department of Justice grant number 2016-DN-BX-K001, and the Tiroler Wissenschaftsfonds (TWF) (UNI-404/1998). We would like to thank Daniel Corach (University of Buenos Aires), Alessandro Achilli (University of Pavia, Italy) and Hovirag Lancioni, (University of Perugia) for providing samples. We would like to thank Tatiana Nogueira and Abdul A. Al-Deib for their technical assistance in obtaining the Brazilian Native American and Libyan samples, respectively, used in this study. We also would like to thank Maiko Takahashi for her technical assistance in this study. The opinions, findings, and conclusions or recommendations expressed in this publication are those of the authors and do not necessarily reflect those of the U.S. Department of Justice.
Publisher Copyright:
© 2019
Keywords
- Forensic genetics
- Haplogroups
- Massively parallel sequencing
- Mitochondrial genome
- MtDNA
- Phylogeny