This study 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.
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. For the current study, just over 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 influence primer binding and, thus, could limit a broad application of this molecular genetic tool. The selected samples derived from various forensically relevant tissue sources were DNA extracted using different methods. The study evaluated sequence concordance and heteroplasmy detection and compared the findings to conventional Sanger sequencing, as well as an orthogonal MPS platform. This article discusses the advantages and limitations of this approach regarding forensic genetic workflow and analytical requirements. (publisher abstract modified)
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