动物造模,荧光定量PCR z-bo 2023-01-18 602

　　Purpose: How to establish an effective method for genotyping Leprdb/+ mouse offspring using TaqMan probe fluorescence quantitative PCR technology?

　　Method: Design a mutation site (rs1801133) to extract 228 mouse offspring of Leprdb mouse tail DNA/+ Lepr gene, a pair of PCR primers and two TaqMan probes? Set the conditions of real-time fluorescent PCR amplification and use SDS software to input SNP sites? Have you passed the obese phenotype verification of a 2-month-old animal to run Hardy? Weinberg balance test?

　　Results: 228 samples were detected by the established TaqMan probe fluorescence quantitative PCR method, among which GG genotype, genotype frequency 0.1929, GT genotype 123, genotype frequency 0.5395, TT genotype 64 and 41 genotype frequencies Is 0.2807? When comparing the typing results of the TaqMan probe fluorescence quantitative PCR method with the obese phenotype results, is the sensitivity 97.56% and the specificity 99.47%?

　　Conclusion: The application of TaqMan probe fluorescence quantitative PCR technology can realize the early typing detection of Leprdb/+ mouse progeny locus.