Microsatellites are widely used markers for multiple purposes in oaks. We describe a complete procedure for cheap DNA extraction and fast microsatellites genotyping by multiplex PCR. 10 loci were selected to form two multiplex kits including three loci that show a high differentiation between Quercus robur and Q. petraea. The loci were tested in three oak species and show a high mean genetic diversity of 0.84. The cumulative exclusion probability for parentage analysis was 0.999977 for single parent and 1.0 for paternity. Finally, the relatively high differentiation coefficient (Gst = 0.04) will facilitate species assignment based on genotypes in oaks