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

Autosomal STR-marker investigations: AmpFISTR MiniFiler (Applied Biosystems)

Y-chromosome STR-marker investigations: AmpFISTR Yfiler kit (Applied Biosystems). The results were evaluated with GeneMapper Software v.4.0 (Applied Biosystems).

Next generation sequencing (NGS)

Budapest-2 laboratory (Dr Erzsébet

Csenák, Dr

Zoltán

Szentirmay)

Analysis of five A-STR markers with sequence capture and subsequent DNA sequencing on the DNA samples of Béla

III

and skeleton 11/52. Out of the investigated 20 A-STR markers, the following 5 were different for the two skeletons, and we wished to decide if these are real differences or artifacts occurring in the degraded DNA samples. The markers were the following: D2S441, D2S1338, D3S1258, D7S820 and D19S433. The process is the following:

a. Creation

of a PCR target panel using Generead DNAseq

Custom Panel v2 kit (Qiagen). A change was necessary in the makeup of the PCR mix due to sample degradation: the reactions final volume is 50pl, in which 25 pl

Kapa Hifi HotStar

Uracil+ enzyme mix (Kapa Biosystems), 10 pl primer mix, 2.5 pl BSA (lmg/ml) and 12.5 pl template can be found. (Note: additional volume of the primer mix / template depends on DNA concentration).

b. Creation

of amplicon library via QIAseq Ultralow Input Library Kit and GeneRead Adapter I Set A 12-plex (Qiagen) kit.

c. Sequencing

via MiSeq Reagent Nano Kit v2 kit on MiSeq

Illumina platform using the following parameters (Table 23).

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