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Massively parallel sequencing-enabled mixture analysis of mitochondrial DNA samples.

Jennifer D Churchill1, Monika Stoljarova2, Jonathan L King3

  • 1Center for Human Identification, University of North Texas Health Science Center, 3500 Camp Bowie Blvd, CBH-250, Fort Worth, TX, 76107, USA. Jennifer.Churchill@unthsc.edu.

International Journal of Legal Medicine
|February 23, 2018
PubMed
Summary

Massively parallel sequencing (MPS) improves mitochondrial DNA analysis for forensic investigations. MPS can identify major contributors in mixed samples, with some success in detecting minor contributors in two-person mixtures.

Keywords:
Ion PGMIon S5Massively parallel sequencingMitochondrial DNAMixtures

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Area of Science:

  • Forensic science
  • Genetics
  • Molecular biology

Background:

  • Mitochondrial DNA (mtDNA) offers valuable forensic information.
  • Interpreting mixed mtDNA samples is challenging for traditional methods.
  • Massively parallel sequencing (MPS) enhances quantitative mtDNA analysis.

Purpose of the Study:

  • To evaluate the effectiveness of MPS in deconvolving mixed mtDNA samples.
  • To assess the capability of MPS in identifying contributors in two- and three-person mixtures.
  • To determine the degree to which MPS can resolve complex mtDNA mixtures.

Main Methods:

  • Preparation of two-person mtDNA mixtures at various ratios (1:1 to 20:1) and three-person mixtures (1:1:1, 5:1:1).
  • Utilized the Precision ID mtDNA Whole Genome Panel and Ion Chef for sample preparation.
  • Sequencing performed on Ion PGM or Ion S5 systems.

Main Results:

  • MPS effectively identified the major contributor in all tested mixtures.
  • Single nucleotide polymorphisms (SNPs) from minor contributors were identified in two-person mixtures up to a 10:1 ratio.
  • Analysis of three-person mixtures was more complex, with complete parsing of mixed haplotypes not achieved.

Conclusions:

  • MPS technologies show promise for interpreting mixed mitochondrial DNA samples in forensic contexts.
  • The effectiveness of MPS in resolving mixtures depends on the number of contributors and their ratios.
  • Further refinement of MPS analysis may improve the deconvolution of complex mtDNA mixtures.