During the last several years, new multi- and single-layered archaeological sites, in which the most ancient Neolithic pottery in the Eastern Europe had been found, were excavated in the region of Lower Volga. Animal bones and organic materials were sampled from these sites for radiocarbon (14C) dating and diet investigations. The evidence from these studies suggests that the first domestic animals in the Lower Volga region appeared in the Cis-Caspian culture of the Early Eneolithic. Lipid analysis of food crusts from pottery allowed the cooked food to be characterized. The detailed chronology from Neolithic (6500–5400 cal BC) to Eneolithic (5300–4700 cal BC) cultures, as well as the diet of these ancient people, were reconstructed.