A panel of ancestry informative markers (AIMs) can be used to describe the genetic components of a population and infer the ancestral origin of a DNA sample. In this study, we selected 30 AIMs from 282 SNPs screened from 30 phenotype-related genes based on the genotyping data of 658 samples from nine populations in the HapMap database. Then,a multiplex assay was developed based on micro-sequencing general chip technologies, and a population allele frequency database was established. This system was utilized to ascertain the origin of subjects from East Asian, European, and African.First, 658 HapMap samples were analyzed using this panel of AIMs, and then 194 unrelated DNA samples from five populations were used for further validation of the system. Finally, population genetic components and individual genetic composition were generated using Structure software, and individual ancestry inferences were made. The 30-AIM assay was well balanced for Hardy-Weinberg equilibrium (P > 0.01), and there was no linkage disequilibrium (l < 0.1). Ancestry component analyses for the 658 HapMap samples and 194 recruited samples were consistent with their known origins. The established panel filtered and developed by the 30 AIMs can be applied to analyze the genetic components of Asian, European,and African populations, as well as individual genetic composition.