show Abstracthide AbstractNew microsatellite markers have been optimized for the development of a sequence based microsatellite genotyping approach. A total of 28 loci were amplified in a multiplexed PCR and sequenced for 156 individuals (from A. alosa, A. fallax and their hybrids) that were repeated to estimate genotyping error. Data were used as a proof of concept of the method and 24 high quality loci were validated. This method has then been applied to genotype additional individuals.