A minimally-invasive method for sampling human petrous bones from the cranial base for ancient DNA analysis.

Ancient DNA (aDNA) research involves invasive and destructive sampling procedures that are often incompatible with anthropological, anatomical, and bioarcheological analyses requiring intact skeletal remains. The osseous labyrinth inside the petrous bone has been shown to yield higher amounts of endogenous DNA than any other skeletal element; however, accessing this labyrinth in cases of a complete or reconstructed skull involves causing major structural damage to the cranial vault or base. Here, we describe a novel cranial base drilling method (CBDM) for accessing the osseous labyrinth from the cranial base that prevents damaging the surrounding cranial features, making it highly complementary to morphological analyses. We assessed this method by comparing the aDNA results from one petrous bone processed using our novel method to its pair, which was processed using established protocols for sampling disarticulated petrous bones. We show a decrease in endogenous DNA and molecular copy numbers when the drilling method is used; however, we also show that this method produces more endogenous DNA and higher copy numbers than any postcranial bone. Our results demonstrate that this minimally-invasive method reduces the loss of genetic data associated with the use of other skeletal elements and enables the combined craniometric and genetic study of individuals with archeological, cultural, and evolutionary value.

[1]  C. Gamba,et al.  Ancient DNA Analysis of 8000 B.C. Near Eastern Farmers Supports an Early Neolithic Pioneer Maritime Colonization of Mainland Europe through Cyprus and the Aegean Islands , 2014, PLoS genetics.

[2]  L. Orlando,et al.  Evolutionary Patterns and Processes: Lessons from Ancient DNA , 2016, Systematic biology.

[3]  M. S. Sørensen,et al.  Estimation of Volume Referent Bone Turnover in the Otic Capsule after Sequential Point Labeling , 2000, The Annals of otology, rhinology, and laryngology.

[4]  R. Durbin,et al.  Sequence analysis Fast and accurate short read alignment with Burrows – Wheeler transform , 2009 .

[5]  Arne Ludwig,et al.  Experimental conditions improving in‐solution target enrichment for ancient DNA , 2017, Molecular ecology resources.

[6]  Swapan Mallick,et al.  Genomic insights into the origin of farming in the ancient Near East , 2016, Nature.

[7]  T. Korneliussen,et al.  Ancient genomics , 2015, Philosophical Transactions of the Royal Society B: Biological Sciences.

[8]  Søren Brunak,et al.  Population genomics of Bronze Age Eurasia , 2015, Nature.

[9]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[10]  M. Slatkin,et al.  Ancient DNA and human history , 2016, Proceedings of the National Academy of Sciences.

[11]  M. Meyer,et al.  Reducing microbial and human contamination in DNA extractions from ancient bones and teeth. , 2015, BioTechniques.

[12]  Improving access to endogenous DNA in ancient bone and teeth , 2015, bioRxiv.

[13]  Bonnie Berger,et al.  Ancient human genomes suggest three ancestral populations for present-day Europeans , 2013, Nature.

[14]  Cristina E. Valdiosera,et al.  Complete mitochondrial genome sequence of a Middle Pleistocene cave bear reconstructed from ultrashort DNA fragments , 2013, Proceedings of the National Academy of Sciences.

[15]  Richard Durbin,et al.  Sequence analysis Fast and accurate short read alignment with Burrows – Wheeler transform , 2009 .

[16]  Eske Willerslev,et al.  Comparing Ancient DNA Preservation in Petrous Bone and Tooth Cementum , 2017, PloS one.

[17]  Swapan Mallick,et al.  Partial uracil–DNA–glycosylase treatment for screening of ancient DNA , 2015, Philosophical Transactions of the Royal Society B: Biological Sciences.

[18]  E. Ekdale,et al.  Comparative Anatomy of the Bony Labyrinth (Inner Ear) of Placental Mammals , 2013, PloS one.

[19]  D. Reich,et al.  The genetic history of Ice Age Europe , 2016, Nature.

[20]  M. S. Sørensen,et al.  The viability and spatial distribution of osteocytes in the human labyrinthine capsule: A quantitative study using vector-based stereology , 2010, Hearing Research.

[21]  K. Veeramah,et al.  Early Neolithic genomes from the eastern Fertile Crescent , 2016, Science.

[22]  M. Meyer,et al.  Single-stranded DNA library preparation for the sequencing of ancient or damaged DNA , 2013, Nature Protocols.

[23]  D. Reich,et al.  Genome-wide patterns of selection in 230 ancient Eurasians , 2015, Nature.

[24]  Matthias Meyer,et al.  Illumina sequencing library preparation for highly multiplexed target capture and sequencing. , 2010, Cold Spring Harbor protocols.

[25]  M. S. Sørensen,et al.  Volume-referent bone turnover estimated from the interlabel area fraction after sequential labeling. , 1998, Bone.

[26]  Marcel Martin Cutadapt removes adapter sequences from high-throughput sequencing reads , 2011 .

[27]  Anders Eriksson,et al.  Upper Palaeolithic genomes reveal deep roots of modern Eurasians , 2015, Nature Communications.

[28]  János Dani,et al.  Genome flux and stasis in a five millennium transect of European prehistory , 2014, Nature Communications.

[29]  N. V. Cramon-Taubadel,et al.  Evolutionary insights into global patterns of human cranial diversity: population history, climatic and dietary effects. , 2014, Journal of anthropological sciences = Rivista di antropologia : JASS.

[30]  A. Eriksson,et al.  The genetics of an early Neolithic pastoralist from the Zagros, Iran , 2016, bioRxiv.

[31]  A. Krogh,et al.  Ancient human genome sequence of an extinct Palaeo-Eskimo , 2010, Nature.

[32]  Kendra Sirak,et al.  Optimal Ancient DNA Yields from the Inner Ear Part of the Human Petrous Bone , 2015, PloS one.

[33]  Mark George Thomas,et al.  Genomic signals of migration and continuity in Britain , 2016 .