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

on MinElute columns with large volume funnels using a QIAvac 24 Plus vacuum system (see above). The extracts were stored at -20°C.

DNA isolation at Budapest-1 laboratory (Dr Judit Olasz,

Dr

Orsolya Csuka)

1. Soaking

the bone samples in a 0.5% NaOCl solution for 15 minutes

2. Three

washes with ultrapure distilled water, overnight drying

3. UV

radiating every side for 10 minutes

4. Pulverization

in a Spex Freezer Mill

5. Decalcination

of 0.15-0.20 g

bone

dust in a solution of 5 ml 0.5 M EDTA (pH8.0) at 4°C for 72 hours (The EDTA solution was changed every 24 hours after prior centrifugation.)

6. DNA

isolation using DNA IQ system (Promega) Kit.

DNA isolation at Budapest-2 Laboratory (Dr Erzsébet

Csernák,

Dr Zoltán

Szentirmay)

1. We

devised the process for sequencing and the creation of the DNA library. The steps are as follows:

2. Preparation

of bone samples (decontamination in order to decrease the amount of inhibitors present):

3. Incubation

of bone samples in a 0.5% NaOCl solution, and UV radiation for 10 minutes.

4. Pulverizing

in a Spex Freezer Mill.

5. Washing

0.15-0.3g bone dust at room temperature in a 1ml 0.5M EDTA (pH8.0) with 15 minutes of incubation.

6. Decalcination

of 0.15-0.3g bone dust at room temperature with adding 1ml 0.5M EDTA for at least 48h (with shaking: 600rpm)

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