Objective To analyze single nucleotide polymorphism(SNP) of inbred mouse genes by a lately established method-single-tube bi-directional allele specific amplification(SB-ASA).Method 16 SNP were detected by the SB-ASA method in 5 inbred mouse strains.We also designed a double-blind experiment and sequencing to validate the reliability of this method,and assessed its influence on the results of PCR system and amplification conditions.Results SB-ASA was successfully used to type 5 inbred mouse strains for 16 SNP loci,and the results were completely consistent with those obtained by sequencing.Five inbred mouse strains were successfully distinguished with a double-blind experiment by just 3 SNP loci.Conclusion SB-ASA can be used to SNP genetic monitoring of inbred mice,and it can be popularized and applied as a new molecular biological method of genetic monitoring.