动物造模,荧光定量PCR z-bio 2022-03-29 286

　　Objective: To establish an efficient method for genotyping the offspring of Leprdb/ + mice using TaqMan probe fluorescence quantitative PCR technology.

　　Methods: The mouse tail DNA of 228 offspring of Leprdb/+ mice was extracted, and one pair of PCR primers and two TaqMan probes were designed for the mutation site of Lepr gene (rs1801133). The software types the SNP loci. It is verified by the obesity phenotype of 2-month-old animals and the Hardy-Weinberg equilibrium test is performed.

　　Results: 228 samples were detected by the established TaqMan probe fluorescence quantitative PCR method, of which 64 samples were GG genotype with a genotype frequency of 0.1929; 123 samples were GT genotype with a genotype frequency of 0.5395; 41 samples were TT genotype The phenotyping frequency was 0.2807. The TaqMan probe fluorescence quantitative PCR method compared the results of phenotyping with obesity, the sensitivity was 97.56%, and the specificity was 99.47%.

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