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Improved DNA Extraction and Illumina Sequencing of DNA Recovered from Aged Rootless Hair Shafts Found in Relics Associated with the Romanov Family.
Federal Bureau of Investigation Laboratory, DNA Support Unit, Quantico, USA.
Linköping University, Faculty of Medicine and Health Sciences. Linköping University, Department of Biomedical and Clinical Sciences, Division of Molecular Medicine and Virology. Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, Sweden.
Federal Bureau of Investigation Laboratory, DNA Support Unit, Quantico, USA.
Federal Bureau of Investigation Laboratory, Trace Evidence Unit, Quantico, USA.
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2022 (English)In: Genes, ISSN 2073-4425, E-ISSN 2073-4425, Vol. 13, no 2, article id 202Article in journal (Refereed) Published
Abstract [en]

This study describes an optimized DNA extraction protocol targeting ultrashort DNA molecules from single rootless hairs. It was applied to the oldest samples available to us: locks of hairs that were found in relics associated with the Romanov family. Published mitochondrial DNA genome sequences of Tsar Nicholas II and his wife, Tsarina Alexandra, made these samples ideal to assess this DNA extraction protocol and evaluate the types of genetic information that can be recovered by sequencing ultrashort fragments. Using this method, the mtGenome of the Tsarina's lineage was identified in hairs that were concealed in a pendant made by Karl Fabergé for Alexandra Feodorovna Romanov. In addition, to determine if the lock originated from more than one individual, two hairs from the locket were extracted independently and converted into Illumina libraries for shotgun sequencing on a NextSeq 500 platform. From these data, autosomal SNPs were analyzed to assess relatedness. The results indicated that the two hairs came from a single individual. Genetic testing of hairs that were found in the second artifact, a framed photograph of Louise of Hesse-Kassel, Queen of Denmark and maternal grandmother of Tsar Nicholas II, revealed that the hair belonged to a woman who shared Tsar Nicholas' maternal lineage, including the well-known point heteroplasmy at position 16169.

Place, publisher, year, edition, pages
2022. Vol. 13, no 2, article id 202
Keywords [en]
Romanov family, ancient DNA, biological sexing, heteroplasmy, hybridization capture, kinship analyses, mitochondrial DNA, next generation sequencing (NGS), single nucleotide polymorphism (SNP)
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Genetics
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URN: urn:nbn:se:liu:diva-193494DOI: 10.3390/genes13020202PubMedID: 35205247OAI: oai:DiVA.org:liu-193494DiVA, id: diva2:1754179
Available from: 2023-05-02 Created: 2023-05-02 Last updated: 2023-05-02

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Tillmar, Andreas

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