Hybridization Enrichment to Improve Forensic Mitochondrial DNA Analysis of Highly Degraded Human Remains

Jennifer M. Young, Denice Higgins, Jeremy J. Austin

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Abstract

Forensic mitochondrial DNA analysis of degraded human remains using PCR-based Sanger sequencing of the control region can be challenging when endogenous DNA is highly fragmented, damaged and at very low concentration. Hybridization enrichment coupled with massively parallel sequencing (MPS) offers an effective alternative for recovering DNA fragments as small as 30 base pairs (bp) from poorly preserved samples. Here, we apply this methodology on a range of degraded human skeletal remains that have previously been analyzed using PCR-based Sanger sequencing with variable success. Our results reaffirm the benefit of targeted enrichment for analysis of degraded remains and highlight the importance of using optimized library preparation and enrichment techniques. We provide an indication of the sequencing depth required to obtain full mtDNA genomes given the complexity of the library and confirm that a second enrichment and/or a very high sequencing effort may be required to obtain full mtDNA genomes for some degraded samples.

Original languageEnglish
Article number450
Number of pages10
JournalFrontiers in Ecology and Evolution
Volume7
DOIs
Publication statusPublished - 20 Nov 2019
Externally publishedYes

Bibliographical note

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

Keywords

  • degraded remains
  • DNA
  • forensic
  • hybridization enrichment
  • massively parallel sequencing
  • mitochondrial genome

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