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

DNA isolation, Göttingen

laboratory

(Verena Seidenberg,

Dr

Susanne Hummel)

We would like to mention in advance, that the DNA isolated from various bone samples had degraded to various degrees, and that the bone samples of Béla

III

and Anne of Antioch were treated with some sort of resin, and this made PCR analyses almost impossible. For this reason, Béla

Ill

’s metatarsal sample was investigated with four special DNA extraction kits (EZ1, QiaVac MinElute Standard, QiaVac MinElute Short, QiaVAc MinElute Organic) and six different DNA extraction methods, out of which method four, five and six were used successfully to isolate PCR DNA templates from various bones.

Initially, the DNA of all ten samples was extracted by two different extraction methods (“QiaVac MinElute Standard” and “EZ1”; see below). Many samples revealed promising amplification results following these extraction methods. However, the samples of Bela III (HU

3B

Mt) and Anne (HU AA Co) in particular were found to contain too many inhibiting substances to enable successful amplification. Therefore, two new extraction methods (“QiaVac MinElute Short” and “QiaVac MinElute Organic”; see below) were developed for optimizing the DNA extraction process.

Sample preparation (for all DNA extraction methods)

The surface of each bone fragment was decontaminated by incubation for 15 min in a commercially available bleach (6% NaOCl) followed by 15 min rinsing in bi-distilled water. The samples were dried overnight at 37°C and then crushed in a steel mortar and powdered in a ball mill (Retsch). 0.25 g

of

bone powder was 225