Abstract: In the Netherlands indigenous Quercus robur L. populations are rare and have been maintained as patches in ancient woodland. For adequate conservation of these populations, information about genetic variation and population structure is necessary. In order to assess the genetic variation and structure of these populations, microsatellite polymorphisms were studied in two autochthonous populations. These two populations differed slightly for their gene diversity, which was as high as was observed for Q. robur populations in France and Germany. For reforestation purposes there is an interest in the genetic variation of a half-sib family harvested from one tree. The gene diversity of the two studied half-sib families - obtained from a forest and an urban area - was similar, but relatively low. This indicates that, for reforestation purposes, seeds should be harvested from many different trees in order to obtain a population with a genetic variation as high as was observed for an autochthonous population.