Objective：To develop a fluorescent labeled multiplex amplification system which is consisted of 5 mini short tandem re－peat（miniSTR） loci（D1S1676，D2S441，D3S4529，D22S1045 and amelogenin） and to analyze the genetic polymorphism of 200 unre－lated Han people in Chongqing. Methods：Primer of D1S1676 was labeled with 5’TAMRA，D2S441 and D3S4529 were labeled with 5’6-FAM，D22S1045 was labeled with 5’HEX and amelogenin was labeled with 5’TET and the multiplex amplification system was established. Data were collected by the 3130 gene analyzer，the length of the amplification fragments was calculated and the genotyp－ing was made using Gene Mapper v3.2.1 software. Results：Every locus of the 5 fluorescent labeled miniSTR multiplex amplification systems can be genotyped clearly. The data of genotyping showed that 9，9，8，9，2 alleles and 103 genotypes were obtained from blood sample of 200 unrelated Han people in Chongqing and the distribution of the genotypes was in accordance with the Hardy-Weinberg equilibrium. The heterozygosis（Ho） and the polymorphism information content（PIC） of the 5 loci in Chongqing Han population were 0.710，0.765，0.730，0.710 and 0.660 041，0.798 329，0.753 177，0.797 888，respectively. The cumulate exclusion probability of pater－nity（CPE） was 0.924 800 4 and the total probability of discrimination power（TPD） was 0.999 955 1. Conclusions：Each locus （D1S1676，D2S441，D3S4529，D22S1045，amelogenin） of the established 5 fluorescent labeled miniSTR multiplex amplification sys－tems can be genotyped accurately and clearly.The genetic data of the 5 loci from Chongqing Han population can be used as basis ma－terials for the study of population genetics and forensic appliance.