Background To conduct phylogeographic or population genetic studies, an adequate number of DNA markers for the focal species are required. Due to severe unavailability of genotype markers of any kind for the species Eurasian minnow (Phoxinus phoxinus L.) and rudd (Scardinius erythrophthalmus L.), we set out to attempt cross-amplification of a set of microsatellite loci from related species. Findings We tested 36 cyprinid microsatellite loci for cross-species amplification in minnow and rudd. Fifteen species-locus combinations produced amplifications in minnow, seven being polymorphic, while 18 combinations amplified in rudd, nine of these being polymorphic. Conclusions The positive cross-species amplifications present potential contributions to the establishment of genetic marker sets for population genetics studies of the two focal species.