Paternity testing was determined in Chinese alligator (Alligator sinensis) clutches during a reproductive season at Xuanzhou Nature Reserve, using five microsatellite loci. DNA from ten mother and offspring clutches was analysed to identify paternal alleles. Three or four paternal alleles were observed among three of ten clutches. These clutches were sired by at least two different males. This present study confirmed the effectiveness of microsatellite DNA markers in detecting multiple paternity within natural populations of Chinese alligator. However, to reduce the confounding effects of mutations and null alleles on allele assignment and to increase power to monitor individual's genetic contribution, we need additional variable genetic markers.