Abstract：A fast and exact sex-specific molecular marker is important in the study of sex control and all-female breeding in half-smooth tongue sole(Cynoglossus semiliaevis). In the present study, a sex-specific co-dominace simple sequence repeat(SSR) marker which had 35 bp difference fragment between the isogeny area of Z and W chromosone was filtrated from 37 same type markers. The selected marker was cloned and sequenced, then the sequence characterized amplified region(SCAR) primer was designed to increase the proportion of the 35 bp defference fragment. Interrelated analyses concluded that 4% agarose gel, 150 V electrophoresis for 25 min were the optimum condition. The transformed SCAR marker could amplificate 134, 134 and 169, 169 bp fragment in super-female、female and male individual, respectively. The SCAR marker was applied in production practice, and 574 physiological male parent half-smooth tongue sole were tested and 100 neo-males were identified and got rid of in less harmful condition which was helpful to increase the female proportion of the offspring. The SCAR marker can be used in accurate test of female, male and super-female individual in laboratory and fast test of male parent half-smooth tongue sole to get rid of neo-male individual under simple and crude environment like fish farm.
