Pisum & Ervilia Tetovac-Made in Early Iron Age Leskovac. Part two. Extraction of Ancient DNA from Charred Seeds from the site of Hissar in South Serbia

Abstract

The extracts were prepared from the samples of 3,200-year-old charred pea and bitter vetch seeds from the site of Hissar near Leskovac, South Serbia, using two different DNA extraction procedures. We used CTAB method with some modifi cation and obtained low quantity of ancient DNA in comparison with the second method used - commercial available kit. After the extraction, a whole genome amplifi cation using Phi29 DNA polymerase was performed. The amplifi ed DNAs were used for PCR reaction using primers for 26S rDNA gene, which is located on the nuclear genome. The single band corresponding to 26S rDNA fragment from modern relatives was obtained. We conclude that DNA from charred pea and vetch seed can be extracted and used for further archaeobotanical analysis at the molecular level.