动物造模,探针荧光定量PCR z-bio 2021-09-14 412

　　Objective: To establish an efficient method for genotyping Leprdb/ + mouse progeny by using TaqMan probe fluorescence quantitative PCR technology.

　　Methods: 228 cases of Leprdb/ + mouse offspring mouse tail DNA were extracted, and 1 pair of PCR primers and 2 TaqMan probes were designed for the mutation site of Lepr gene (rs1801133). Set conditions for real-time fluorescence PCR amplification, using SDS The software classifies the SNP loci. Passed the obesity phenotype verification of 2-month-old animals and performed the Hardy-Weinberg balance test.

　　Results: The established TaqMan probe fluorescence quantitative PCR method was used to detect 228 samples, including 64 GG genotypes with a genotype frequency of 0.1929; GT genotypes with a genotype frequency of 0.5395; TT genotypes with 41 genes The type frequency is 0.2807. Compared with the typing results of the TaqMan probe fluorescence quantitative PCR method and the obesity phenotyping results, the sensitivity is 97.56% and the specificity is 99.47%.

　　Conclusion: The application of TaqMan probe fluorescence quantitative PCR technology can realize the early typing detection of Lepr db/ + mouse progeny gene loci, the method is simple and efficient.