The Fosse Ardeatine massacre was a mass execution carried out in Rome on March 24, 1944 by Nazi German occupation troops during the Second World War as a reprisal for a partisan attack conducted on the previous day in central Rome. The 335 civilians were taken to the "Cave Ardeatine" and they were shot. Only 323 corpses out of 335 have been identified. The aim of this work is the genetic and anthropological analysis of the remains exhumed from grave number 329 of Fosse Ardeatine's Shrine to assess their identity. So far, such remains have been supposed to belong to MM but mitochondrial analysis excluded a biological relationship to two living maternal relatives. Our analysis indicated that remains recovered in grave number 329 do not belong to MM. This result suggests that genetic analysis of the remains should be also applied to the other 12 unknown corpses to elucidate their identity.