Radiocarbon dating
Radiocarbon dating of the Mokp individual (UCIAMS210906: 2205 ± 20 BP, 364–197 International Radiocarbon Calibration Curve (2020 version, IntCal20) calibrated years bce) was carried out at the Keck Laboratory, University of California, Irvine, following the methodology described by Librado et al.50.
Sex and age-at-death estimations of the human remains
Age-at-death determination methods rely on a variety of skeletal indicators, including stages of auricular surface for adults51,52, stages of iliac crest or sternal end of the clavicle fusion, measurement of long bones for immature individuals53,54 and dental eruption sequences55,56. Biological sex is on the basis of genetic data, especially the so-called Ry ratio (Y to Y + X sequence coverage)57 (Supplementary Table 1a).
DNA extraction
Samples were processed in the clean laboratory facilities at the Centre for Anthropobiology and Genomics of Toulouse (CAGT), University of Toulouse, or at the Centre for GeoGenetics (CGG), University of Copenhagen, following ancient DNA procedures (Supplementary Information section 2.2).
Bone and tooth samples
After gentle surface abrasion, a portion of the dense part of the bone samples was collected using a diamond wheel (PROXXON or ARGOFILE instruments). For tooth samples, the cementum was isolated as recommended by Damgaard et al.58. The samples were either crushed into smaller fragments using a manual mortar or cutting pliers, or pulverized using a Retsch MM200 instrument and then placed in 5-ml Eppendorf LoBind tubes. DNA was extracted following a silica-column-based method, as described by Librado et al.59, without bleach pretreatment (Supplementary Information section 2.2).
Calculus samples
Calculus samples were isolated, as described by Sabin and Yates60. Samples labelled as ‘Name_C’ in Extended Data Fig. 4a (for example, Eletchei3_C_C_P4) were extracted for DNA following a protocol similar to that used for bones and cementum, except that no 1-h predigestion was performed and the digestion volume was limited to 1 ml. Samples labelled as ‘Name_CE’ (for example, Eletchei3_CE_C_P4) were subjected to an overnight digestion at 50 °C in 555 µl of a buffer consisting of 0.45 M EDTA, 1.8 mg ml−1 of proteinase K and 9 mM dithiothreitol. The supernatant was further purified on a QIAGEN MinElute column and eluted in 40-µl sterile water.
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