Mitochondrial DNA sequencing of ancient samples from Polizzello (Sicily)

To test the efficiency of our computational pipeline for mitochondrial DNA (mtDNA) reconstruction and variant detection, we analyzed 30 Iron Age samples from Polizzello (Sicily), an archaeological site in the heart of Sikania dated at IX-VII centuries B.C. Molecular analysis was carried out in the Laboratory of Anthropology and Paleogenetics at University of Florence, on petrous bones where endogenous ancient DNA is better preserved than in other anatomical districts, following strict guidelines to prevent modern contamination during all experimental steps. After library preparation following a custom double-indexing protocol optimized for ancient samples, mtDNA molecules were selected using hybrid-capture target enrichment strategy. Enriched libraries were then sequenced on Illumina NovaSeq 6000 platform.