The greater horseshoe bat, Rhinolophus ferrumequinum (Schreber, 1774), is one of the most distributed bat species in Turkey. This study is based on statistical evaluation of the cranial measurements of 165 specimens for determining whether a clinal increase in the species throughout the country. In addition, conventional stained karyotype along with C- and Ag-NOR banded chromosomes of the specimens were studied. No clinal increase in size is encountered from west to east in the greater horseshoe bat. On the contrary, Southeastern Anatolia, Eastern Anatolia, Central Anatolia, western and central Black Sea regions form a separate cluster, while the Mediterranean, Aegean and Marmara regions form another one with regard to cranial measurements. In addition, the populations do not differ in terms of their karyotype and banded chromosomes.