Conservation of aquaculture or characteristic fish requires knowledge of genetic variation within and between breeds. Genetic marker are molecular labels which identify genes and are valuable to the animal industry as tools of animal selection. In this study, Microsatellite DNA techniques will chose for identify the genetic diversity within and among different populations for Culter spp. The Erythroculter ilishaeformis is a economic species to freshwater streams or lake in Taiwan, such as Sun-Moon Lake. Microsatellite loci were characterized in the E. ilishaeformis random clones from small genomic library using (GT)15 and (CT)15 prob. Primers for DNA amplification using PCR were designed and synthesized for 5 loci in this research. Developed microsatellite primers should prove useful for population studies of the E. ilishaeformis.