title: Massively parallel sequencing of complete mitochondrial genomes from hair shaft samples.
abstract: Though shed hairs are one of the most commonly encountered evidence types, they are among the most limited in terms of DNA quantity and quality. As a result, DNA testing has historically focused on the recovery of just about 600 base pairs of the mitochondrial DNA control region. Here, we describe our success in recovering complete mitochondrial genome (mtGenome) data (∼16,569bp) from single shed hairs. By employing massively parallel sequencing (MPS), we demonstrate that particular hair samples yield DNA sufficient in quantity and quality to produce 2-3kb mtGenome amplicons and that entire mtGenome data can be recovered from hair extracts even without PCR enrichment. Most importantly, we describe a small amplicon multiplex assay comprised of sixty-two primer sets that can be routinely applied to the compromised hair samples typically encountered in forensic casework. In all samples tested here, the MPS data recovered using any one of the three methods were consistent with the control Sanger sequence data developed from high quality known specimens. Given the recently demonstrated value of complete mtGenome data in terms of discrimination power among randomly sampled individuals, the possibility of recovering mtGenome data from the most compromised and limited evidentiary material is likely to vastly increase the utility of mtDNA testing for hair evidence.
authors.affiliation: Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria; Penn State Eberly College of Science, University Park, PA, USA. Electronic address: walther.parson@i-med.ac.at.
authors.forename: Walther
authors.initials: W
authors.lastname: Parson
authors.affiliation: Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Huber
authors.affiliation: FBI Laboratory, Quantico, VA, USA.
authors.forename: Lilliana
authors.initials: L
authors.lastname: Moreno
authors.affiliation: Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: Maria-Bernadette
authors.initials: MB
authors.lastname: Madel
authors.affiliation: FBI Laboratory, Quantico, VA, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Brandhagen
authors.affiliation: Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: Simone
authors.initials: S
authors.lastname: Nagl
authors.affiliation: Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: Catarina
authors.initials: C
authors.lastname: Xavier
authors.affiliation: Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria.
authors.forename: Mayra
authors.initials: M
authors.lastname: Eduardoff
authors.affiliation: FBI Laboratory, Quantico, VA, USA.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Callaghan
authors.affiliation: FBI Laboratory, Quantico, VA, USA. Electronic address: jodi.irwin@ic.fbi.gov.
authors.forename: Jodi A
authors.initials: JA
authors.lastname: Irwin
mesh_terms: D004272:DNA, Mitochondrial / Q000235:genetics*; D053803:Forensic Genetics; D054629:Genome, Mitochondrial; D006197:Hair / Q000737:chemistry*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans
chemical_list: D004272:DNA, Mitochondrial
keywords: Forensic science; Heteroplasmy; Next generation sequencing; mtDNA; mtGenomes