Genetic studies between related species can be used to determine their genetic variability and their use in the development of new varieties. Aegilops tauschii and Triticum durum are two important species, used as genetic resources in the wheat breeding program at Embrapa. The objective of the present study was to evaluate the genetic similarity between 14 accesses of Ae. tauschii, five of T. durum and nine of the resulting hybrids originated from the crossing between the two species. Total genomic DNA was extracted by CTAB method, from young leaves, as protocol used at Embrapa Trigo. DNA was quantified using a spectrophotometer and it was diluted to 50 ng/ul. PCR amplification cycles were performed as follow: 94ºC/3 minutes, 45 cycles of 94ºC/1 minute, 60ºC/1 minute (primers annealing temperatures were 50ºC, 55ºC or 60ºC), 72ºC/2 minutes and finally 72ºC/10 minutes. The amplification products were separated on 2% agarose gel and visualized by ethidium bromide staining method. The genetic similarity was determined using NTSYS-pc software and a dendrogram was generated using UPGMA cluster analysis. The dendrogram derived from UPGMA analysis were based on 15 polymorphic primers and generated three distinct groups: a group formed by Ae. Tauschii, another one for T. durum and the third for their hybrids. Moreover, intra-specific variability in each group was detected, and the highest similarity coefficient was 0,92.