In Tunisia, the fig (Ficus carica L.) is one of the fruit species characterized by large adaptative potentialities to various ecological areas since it is growing in numerous environmental and soil conditions. For instance fig is, after date-palms, the main fruit crop cultivated in the oases. Tunisian fig germplasm is characterized by a high genetic diversity characterized by a large number of cultivars. However, only a few of them, chosen for their fruit traits (pulpy and sweetly, high fruit yields, etc) have spread widely in the different groves. Therefore, this phytogenetic resource is threatened by severe genetic erosion due to either their vulnerability to biotic and abiotic pressures and intensive urbanisation. Recently, prospecting and collecting actions have been initiated and led to the identification of more than 50 ecotypes. These are ex situ maintained in four collections at the "Institut des Regions Arides" of Medenine, the "Centre de Recherches Phoenicicoles" of Degache, the "Commissariat Regional du Developpement Agricole" of Gafsa and the "Ecole Superieure d'Horticulture et d'Elevage" of Chott Mariam. It should be stressed that the actual number of cultivars is at present difficult to estimate since synonymy constitutes a source of error in varietal identification. Multiple studies have been carried out describing the genetic diversity in this crop and aimed at the molecular characterization of the local germplasm. Here we report the development of the random amplified microsatellite polymorphism (RAMPO), a polymerase chain reaction method, in order to enlarge the number of molecular markers reliable in the characterization of Tunisian fig cultivars. Data proved that this procedure constitutes an efficient tool to provide molecular markers suitable for assessment of molecular polymorphisms in this crop. The feasibility and the opportunity of the RAMPO technique are also discussed in relation to the characterization of the local rig germplasm.