High-quality and high-throughput massively parallel sequencing of the human mitochondrial genome using the Illumina MiSeq

Jonathan L King; Bobby L LaRue; Nicole M Novroski; Monika Stoljarova; Seung Bum Seo; Xiangpei Zeng; David H Warshauer; Carey P Davis; Walther Parson; Antti Sajantila; Bruce Budowle

doi:10.1016/j.fsigen.2014.06.001

High-quality and high-throughput massively parallel sequencing of the human mitochondrial genome using the Illumina MiSeq

Forensic Sci Int Genet. 2014 Sep:12:128-35. doi: 10.1016/j.fsigen.2014.06.001. Epub 2014 Jun 7.

Affiliations

¹ Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, TX 76107, USA. Electronic address: Jonathan.King@unthsc.edu.
² Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, TX 76107, USA.
³ Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria; Penn State Eberly College of Science, University Park, PA, USA.
⁴ Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, TX 76107, USA; Department of Forensic Medicine, Hjelt Institute, P.O. Box 40, 00014 University of Helsinki, Helsinki, Finland.
⁵ Institute of Applied Genetics, Department of Molecular and Medical Genetics, University of North Texas Health Science Center, 3500 Camp Bowie Blvd., Fort Worth, TX 76107, USA; Center of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah, Saudi Arabia.

PMID: 24973578
DOI: 10.1016/j.fsigen.2014.06.001

Abstract

Mitochondrial DNA typing in forensic genetics has been performed traditionally using Sanger-type sequencing. Consequently sequencing of a relatively-large target such as the mitochondrial genome (mtGenome) is laborious and time consuming. Thus, sequencing typically focuses on the control region due to its high concentration of variation. Massively parallel sequencing (MPS) has become more accessible in recent years allowing for high-throughput processing of large target areas. In this study, Nextera(®) XT DNA Sample Preparation Kit and the Illumina MiSeq™ were utilized to generate quality whole genome mitochondrial haplotypes from 283 individuals in a both cost-effective and rapid manner. Results showed that haplotypes can be generated at a high depth of coverage with limited strand bias. The distribution of variants across the mitochondrial genome was described and demonstrated greater variation within the coding region than the non-coding region. Haplotype and haplogroup diversity were described with respect to whole mtGenome and HVI/HVII. An overall increase in haplotype or genetic diversity and random match probability, as well as better haplogroup assignment demonstrates that MPS of the mtGenome using the Illumina MiSeq system is a viable and reliable methodology.

Keywords: Discrimination power; Haplogroup assignment; Illumina MiSeq; Massively parallel sequencing (MPS); Mitochondrial genome (mtGenome); Mitochondrial haplotype.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA, Mitochondrial / genetics*
Genome, Mitochondrial*
Haplotypes
High-Throughput Nucleotide Sequencing / methods*
Humans

Substances

DNA, Mitochondrial