Microsatellites are playing an important role in paternity assignment of animals. Given the cost and effort, it would be optimal to develop a minimal microsatellite marker set for paternity testing. This study was the first to assess paternity in a captive colony of rhesus macaques (Macaca mulatta) from the Chinese province of Anhui using 10 polymorphic microsatellites. Results indicated that if at least 6 loci were genotyped, the probability of paternity assignment success was nearly 100%. Our results provide a panel of 6 markers that is effective for assessing paternity of subspecies M. m. siamica of Anhui origin.