Tigridia pavonia (L.f.) DC. is one of the important phytogenetic resources of México. This species is used as ornamental, food and medicinal purposes. Despite its ornamental and economic potential, there is little information about the genetic variability. In this study, randomly amplified polymorphic DNA (RAPD) primers of 10, 15 and 20 bases were used to assess the level of genetic variation among nine botanical varieties of Tigridia pavoniacollected in three localities within State of México. The total number fragments, polymorphic fragments, percentage of polymorphism and resolving power were greater for 15 base (55, 52, 94.5 and 5, respectively) and 20 base (47, 45, 95.7 and 3.8, respectively), in comparison with those obtained from 10 base primers (44, 41, 93.1 and 3.6, respectively).Resultsshowed the major effectiveness of 15 and 20 bases RAPD primers in the genetic differentiation of varieties as compared to 10 bases RAPD primers. The dendrograms based on un-weighted pair group method arithmetic average (UPGMA) analysis of the 10, 15, 20 and the pooled (10, 15 and 20) bases RAPD data were consistent in the clustering varieties, grouping them in two main clusters.