Abstract:The method of cross-amplification can be used to obtain microsatellite marker quickly and effectively. This study amplified microsatellite loci of Macroclemys temminckii using published microsatellite DNA primers from related species. In M.temminckii, 8 microsatellite loci which had polymorphism were found in 69 microsatellite markers. By analyzing PCR amplification products, a total of 30 alleles were detected, and the average number of alleles was 3.75. The observed heterozygosity (Ho) ranged from 0.041 7 to 0.954 5, and expected heterozygosity (HE) was 0.041 7 to 0.811 8. PIC value was from 0.040 0 to 0.759 2, averaging 0.423 2. After chi-square test, some loci were found to correspond to Hardy-Weinberg equilibrium. In general, these loci are good molecular markers that can be used for studying genetic structure for M.temminckii.