Objective Analysis of the genetic structure stability of Brandt's vole (lasiopodomys brandtii) in closed colony using microsatellite marker technology. Methods Genomic DNA was extracted from tail tip using high-concentration-salt precipitation methods. Marked with fluorescent tags(Fam), 7 microsatellite primers were filtered out by PCR, and the DNA structure of three consecutive generations of Brandt's vole was analyzed by microsatellite marker. Results By the analysis of the average heterozygosity and polymorphism information content, Brandt's vole populations maintaineda closed group of qualified genetic structure. Conclusions The present results show that the closed group of Brandt's vole species in our laboratory maintain a stable genetic structure.