Abstract: In this study, we established a technique for the parentage assignment in Asian Yellow Pond Turtles(Mauremys mutica) based on two multiplex PCR panels. The primer ratio, the concentration of the fluorescent joint, the annealing temperature and the number of cycles were optimized in this technique. Each multiplex PCR panel had eight loci, 10 of which were developed in our lab and the other 6 were chosen from published literatures. With the multiplex PCR tool we performed the genotyping and the genetic diversity analysis on 428 individuals using ABl3I30 genetic an-alyzer and Cervus 3.0 software. The results showed that the average number of alleles was 14.190; the average poly-morphism information content(PIC) was 0.748; the average observed heterozygosity(Ho) and expected heterozygosity(He) were 0.687 and 0.771 respectively. The parentage assignment was performed on 89 candidate female parents and 296 offspring. Without information on the male parent, the identification rate was 87%. The numbers of offspring of different female parents were remarkably various, ranging from 1 to 12. This provided the material foundation for the selective breeding. This technique could be a highly efficient tool for the genetic diversity analysis, the marker-assisted family identification and management, and the selective breeding of Asian Yellow Pond Turtles.