Ancient DNA Methods Improve Forensic DNA Profiling of Korean War and World War II Unknowns.

Elena I Zavala,Kerriann K Meyers,Jennifer Daniels-Higginbotham,Birgit Nickel,Charla Marshall,Timothy P Mcmahon,Julia Richter,Gregory E Berg,Suzanne Barrit-Ross,Ayinuer Aximu-Petri,Jacqueline Tyler Thomas,Kimberly Sturk-Andreaggi,Matthias Meyer

doi:10.3390/genes13010129

Abstract

The integration of massively parallel sequencing (MPS) technology into forensic casework has been of particular benefit to the identification of unknown military service members. However, highly degraded or chemically treated skeletal remains often fail to provide usable DNA profiles, even with sensitive mitochondrial (mt) DNA capture and MPS methods. In parallel, the ancient DNA field has developed workflows specifically for degraded DNA, resulting in the successful recovery of nuclear DNA and mtDNA from skeletal remains as well as sediment over 100,000 years old. In this study we use a set of disinterred skeletal remains from the Korean War and World War II to test if ancient DNA extraction and library preparation methods improve forensic DNA profiling. We identified an ancient DNA extraction protocol that resulted in the recovery of significantly more human mtDNA fragments than protocols previously used in casework. In addition, utilizing single-stranded rather than double-stranded library preparation resulted in increased attainment of reportable mtDNA profiles. This study emphasizes that the combination of ancient DNA extraction and library preparation methods evaluated here increases the success rate of DNA profiling, and likelihood of identifying historical remains.

Highlights

Licensee MDPI, Basel, Switzerland.Massively parallel sequencing (MPS), known as next-generation sequencing (NGS), has only recently begun integration into the forensic field
Six extraction/repair conditions were tested in total (Table 2); the two ancient DNA extractions were from the same original powder sample
We found that the percent of sequences that mapped to the human reference genome for about half of the historical remains fell within two standard deviations of the mean for the ancient samples (Figure 3A)

Summary

Introduction

Licensee MDPI, Basel, Switzerland.Massively parallel sequencing (MPS), known as next-generation sequencing (NGS), has only recently begun integration into the forensic field. MPS was first accepted in a court case in the Netherlands in 2019 [1]. While more challenging cases involving historical skeletal remains, including historical figures, unmarked graves, and unidentified war victims, have seen some advances in DNA profiling with MPS, success rates are still low [4,5,6]. This may be due to the fact that successful forensic DNA profiling typically requires relatively large

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Results

Conclusion