P040 – A Genetic Investigation of a Medieval Slovenian Cemetery Using Next-Generation Sequencing

11:13

Wednesday August 19^th

Station 08

Duration: 12 minutes

03. Human identification

Madalynn Martino

In this project, DNA testing was performed using forensic methods on 31 petrous bones representing 31 individuals found in burials dated to late 8th, early 11th century Slovenia. Remains were discovered next to The Church of the Assumption of Mary in the town of Muljava during three excavations (1988, 2012, and 2016). This project leveraged next generation sequencing of lineage markers and autosomal SNPs to predict sex, ancestry, phenotype and familial relationships of the individuals.

DNA was extracted using a non-organic method developed to recover small fragments and then the DNA was repaired using a USER treatment. Libraries were created, followed by enrichment for different targets using three separate hybridization capture panels. One panel was used to target the whole mitochondrial genome (mitogenome), while the FORCE and 95K SNP panels were utilized to target nuclear SNPs. Sequence data were generated on Illumina systems and analyzed using the CLC Genomics Workbench for the mitogenome panel or the Parabon Fx Forensic Analysis Platform for the SNP panels.

Reportable data were generated for 28 of the 31 individuals, with no data generated for 3 individuals due to insufficient human DNA content. Full mitogenome profiles were obtained with an average coverage of 6843X across the samples, with 24 unique haplotypes identified. The 28 samples also obtained an average recovery of 4586 1X SNPs for the FORCE panel and 74730 1X SNPs for the 95K panel. These SNP recoveries were sufficient to provide a prediction of sex, Y haplogroup, continental ancestry, and phenotype. Furthermore, kinship inferences out to 5th degree were obtained with strong statistical support, revealing relatedness within the burials of medieval Slavs. Ultimately, this project successfully applied forensic methods to ancient DNA, unveiling detailed genetic and familial insights into the early medieval Slavic community.

Disclaimer: The opinions or assertions presented hereafter are the private views of the speaker(s) and should not be construed as official or as reflecting the views of the Department of Defense, its branches, the Defense Health Agency, or the Armed Forces Medical Examiner System. Any mention of commercial products was done for scientific transparency and should not be viewed as endorsement of the product or manufacturer.

Authors

Madalynn Martino (Armed Forces Medical Examiner Systems, United States of America)
Kimberly Sturk-Andreaggi (Armed Forces Medical Examiner Systems, United States of America)
Courtney Cavagnino (Armed Forces Medical Examiner Systems, United States of America)
Matija Črešna (Centre for Interdisciplinary Research in Archaeology, Slovenia)
Tamara Leskovar (Centre for Interdisciplinary Research in Archaeology, Slovenia)
Saso Porenta (Terarhis d.o.o. and Primož Stergar s.p., Slovenia)
Jayne-Leigh Thomas (Indiana University, United States of America)
Charla Marshall (Pennsylvania State University, United States of America)

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P040 – A Genetic Investigation of a Medieval Slovenian Cemetery Using Next-Generation Sequencing

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