Objective To explore the application of multiplex PCR method in the detection of microsatellite in mice. Methods\ Four pairs of specific primers were selected to perform multiplex DNA amplification in ten inbred stains of mice. Results\ Duplex DNA amplification got two expected bands under the same condition of the PCR reaction, but triplex DNA amplification did not get three expected bands.Conclusion\ The duplex DNA amplification is a fast and economical method to analyse genotype of inbred mice.