Miklós Kásler - Zoltán Szentirmay (szerk.): Identifying the Árpád Dynasty Skeletons Interred in the Matthias Church. Applying data from historical, archaeological, anthropological, radiological, morphological, radiocarbon dating and genetic research (Budapest, 2021)
Investigated bone samples and methods
incubated rotating with 3,900 pl EDTA (0.5 M; pH 8) and 100 pl Proteinase K at 37°C for 18h. EZ1 Subsequently, further 50 pl of Proteinase K was added and the samples were rotated for another 2 h at 56°C. Subsequently, 50 pl of SDS was added followed by incubation for 5 min at 65°C. The lysate was centrifuged for 3 min at 3,300 ref. The supernatant was transferred to Amicon® Ultra-4 Centrifugal Filter Devices 30 K (Millipore) and concentrated to approx. 250 ml by centrifugation at 5,000 ref. The remaining lysate was purified in the BioRobot EZ1 using the Trace Protocol on the Forensic Card and the EZ1 DNA Tissue Kit (all components, soft- and hardware Qiagen). The elution volume was 50 pl. The extracts were stored at -20°C. Qia Vac MinElute Standard The duration of the EDTA/Proteinase K incubation was 18 h. Subsequently, a further 50 pl of Proteinase K was added and the samples were rotated for another 2 h at 56°C. Then 50 pl of SDS was added followed by incubation for 5 min at 65°C. The lysate was centrifuged for 3 min at 3,300 ref. The supernatant was mixed with 16 ml of PB buffer (Qiagen) and 100 pl sodium acetate buffer (pH 5.2), centrifuged for 3 min at 3,300 ref again and transferred to MinElute columns with large volume funnels on a QIAvac 24 Plus vacuum system (both Qiagen). The lysate was pulled through by vacuum, followed by three washing steps with 700 pl PE buffer waiting each time for 5 min before 226
