Abstract: Numerous studies have found that genes such as LDHA, F-KER, DRD4, MSTN, and CRY1 in racing pigeons exhibit single nucleotide polymorphisms（SNPs）that are associated with homing ability, making them potential SNP markers for molecular-assisted breeding. Current detection methods are still predominantly based on traditional RFLP, and the application of Sanger sequencing has not been reported yet. This study aims to establish a detection method based on Sanger sequencing technology to identify polymorphic sites in these five genes. The results demonstrate that specific primers for the SNPs of the LDHA, F-KER, DRD4, MSTN, and CRY1 genes in racing pigeons were designed, and the amplified PCR products yielded single, specific bands. Furthermore, using these primers, amplification and Sanger sequencing were performed on feather DNA samples from 60 racing pigeons, successfully obtaining polymorphic typing results for each gene locus. Finally, the distribution frequency of various genotypes among these 60 racing pigeons was statistically analyzed, showing that the frequencies of the relevant rare genotypes were all below 5%, consistent with the frequencies reported in foreign racing pigeon populations. The Sanger sequencing analysis method established in this study for SNP typing of racing pigeon performance-related genes holds significant practical value for breeding superior genotypes of racing pigeons.