Working with mitochondrial DNA from highly degraded samples is challenging. We present a whole mitogenome Illumina-based sequencing method suitable for highly degraded samples. The method makes use of double-stranded library preparation with hybridization-based target enrichment. The aim of the study was to implement a new user-friendly method for analysing many ancient DNA samples at low cost. The method combines the Swift 2S™ Turbo library preparation kit and xGen® panel for mitogenome enrichment. Swift allows to use low input of aDNA and own adapters and primers, handles inhibitors well, and has only two purification steps. xGen is straightforward to use and is able to leverage already pooled libraries. Given the ancient DNA is more challenging to work with, the protocol was developed with several improvements, especially multiplying DNA input in case of low concentration DNA extractions followed by AMPure® beads size selection and real-time pre-capture PCR monitoring in order to avoid cycle-optimization step. Nine out of eleven analysed samples successfully retrieved mitogenomes. Hence, our method provides an effective analysis of whole mtDNA, and has proven to be fast, cost-effective, straightforward, with utilisation in population-wide research of burial sites.

CF Genomics CEITEC Masaryk University Kamenice 753 5 Brno 62500 Czech Republic

Institute for Archaeological Heritage Kaloudova 1321 30 Brno 61400 Czech Republic

Laboratory of Biological and Molecular Anthropology Department of Experimental Biology Faculty of Science Masaryk University Kamenice 753 5 Brno 62500 Czech Republic

Masaryk Museum Zamecke Nam 27 9 Hodonin 69501 Czech Republic

Citace poskytuje Crossref.org

Extraction Protocol for Parallel Analysis of Proteins and DNA from Ancient Teeth and Dental Calculus