Extraction and analysis of ancient DNA from human remains of Masjede Kabood burial site

Extraction and analysis of DNA from ancient remains has numerous applications in archeology and molecular evolution. However, it has become obvious that ancient DNA (aDNA) can be easily contaminated with modern DNA, so it is crucial to detect contamination and to distinguish contaminant from authentic results. In the present study, we report the successful extraction and amplification of aDNA from 3000-3500 yearold human remains excavated from Masjede kabood (Tabriz, North-West of Iran) burial site. To test the authenticity of the extracted aDNA, we have developed a nested PCR/restriction enzyme digestion method for molecular sex determination of the skeletal remains, which their gender was known based on their morphology and belongings (Crown, Sword, Bracelet etc.). A simple and effective modified ethanol precipitation-based protocol was used for DNA extraction from 35 human skeletal remains. A segment of Homologous Amelogenin Gene (AMG), which has different alleles on X and Y-chromosomes, was amplified and analyzed. The obtained data were compared with anthropometric reports as a control for the rate of precision in aDNA analysis. The results showed that reliable aDNA can be extracted and amplified from archeological remains. The presented sex determination procedure could also be used as a reliable control for testing the authenticity of aDNA results.