Our investigation was based on a molecular study of the genetic relationships among raspberry genotypes collected around selected medieval castles, Carthusian monasteries and nearby villages. We assumed that the hypothetical medieval raspberry genotypes could be traced to isolated medieval settlements that used to be highly prosperous during the feudal era but were later abandoned. Some of these genotypes could have survived in natural conditions without seed multiplication for at least three centuries. The molecular genetic analysis was based on microsatellite data. A total of 155 alleles were detected at 18 microsatellite loci. The clustering method grouped the analysed genotypes into seven main clusters. The analyses indicated that the most probable medieval genotypes had been collected around the ruins of two abandoned Carthusian monasteries: Zice and Jurkloster. They were morphologically very similar, vigorous and primitive but obviously not wild genotypes. The plants could be more than 2.3 m high, the canes were medium waxy, the lower and upper parts of the canes were covered by sparse short spines, the mid part was more or less completely smooth, the fully developed leaves were 15–25 cm long and the inflorescences were loose. In addition, the flowers were relatively small, the fruit setting was poor and the fruits were small, ovoid to conical and more aromatic than those of modern cultivars.