Hypervariable minisatellite DNA probes 33.15 and 33.6, originally developed for studies in human populations, were used to study genetic variation in chamois (Rupicapra rupicapra). The mean number of bands per individual was 25 for probe 33.15 and 15 for probe 33.6. The average band frequency was 0.33 for both probes. The mean similarity was 0.44, greater than that reported for human and natural populations and close to values found in domestic populations of mammals. This lack of variability could be related to the bottleneck suffered by the population due to large-scale hunting after the Spanish Civil War. Levels of variability are high compared with variability at the level of protein markers, so the use of minisatellite DNA is recommended for future population studies in this species. We did not find large genetic differences between subpopulations, indicating that the population is genetically homogeneous.