International Journal of Legal Medicine

, Volume 132, Issue 5, pp 1263–1272 | Cite as

Massively parallel sequencing-enabled mixture analysis of mitochondrial DNA samples

  • Jennifer D. ChurchillEmail author
  • Monika Stoljarova
  • Jonathan L. King
  • Bruce Budowle
Original Article


The mitochondrial genome has a number of characteristics that provide useful information to forensic investigations. Massively parallel sequencing (MPS) technologies offer improvements to the quantitative analysis of the mitochondrial genome, specifically the interpretation of mixed mitochondrial samples. Two-person mixtures with nuclear DNA ratios of 1:1, 5:1, 10:1, and 20:1 of individuals from different and similar phylogenetic backgrounds and three-person mixtures with nuclear DNA ratios of 1:1:1 and 5:1:1 were prepared using the Precision ID mtDNA Whole Genome Panel and Ion Chef, and sequenced on the Ion PGM or Ion S5 sequencer (Thermo Fisher Scientific, Waltham, MA, USA). These data were used to evaluate whether and to what degree MPS mixtures could be deconvolved. Analysis was effective in identifying the major contributor in each instance, while SNPs from the minor contributor’s haplotype only were identified in the 1:1, 5:1, and 10:1 two-person mixtures. While the major contributor was identified from the 5:1:1 mixture, analysis of the three-person mixtures was more complex, and the mixed haplotypes could not be completely parsed. These results indicate that mixed mitochondrial DNA samples may be interpreted with the use of MPS technologies.


Mitochondrial DNA Mixtures Massively parallel sequencing Ion S5 Ion PGM 



We would like to thank Thermo Fisher Scientific for providing reagents and technical support necessary to perform this study.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Center for Human IdentificationUniversity of North Texas Health Science CenterFort WorthUSA
  2. 2.Department of Chemistry and BiotechnologyTallinn University of TechnologyTallinnEstonia
  3. 3.Center of Excellence in Genomic Medicine Research (CEGMR)King Abdulaziz UniversityJeddahSaudi Arabia

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