Methods for the analysis of mitochondrial DNA

The use of mitochondrial DNA (mtDNA) profiling as a tool for forming a link between an individual and a crime sample is a well‐established forensic biology technique. Though nuclear DNA typing is considered the gold standard method, certain evidence types have limited nuclear DNA available, such as telogen hairs and ancient or degraded remains, for which mtDNA analysis may be required. Traditionally mtDNA typing has been carried out using Sanger type sequencing, which is labor, time, and cost intensive. This has restricted the analysis of the mitochondrial genome to the control region, limiting the discrimination power of the technique. The introduction of massively parallel sequencing (MPS) into forensic laboratories has brought about the ability to analyze the whole mitochondrial genome of forensically relevant samples. The use of MPS for mtDNA analysis has resulted in an increase in the range of methods available for analysis, including: midi‐ and mini‐sized amplification strategies for whole genome analysis of casework‐type samples; whole genome single nucleotide polymorphism (SNP) multiplexes; and DNA capture methods allowing the analysis of highly degraded remains. The resulting increase in discriminatory power and the continued progression of the technology and associated methods increase its appeal as a DNA analysis method and may facilitate its implementation as a routinely used forensic tool. Here, we give an overview of current laboratory practices for the analysis of mtDNA using MPS. This article is categorized under: Forensic Biology > Haploid Markers Forensic Biology > Forensic DNA Technologies