Genotyping of the offsprings of Lepr db/ + mice by TaqMan probe fluorescence quantitative PCR
Received:September 04, 2017  
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DOI:10.3969/j. issn. 1005 -4847. 2018. 02. 011
KeyWord:TaqMan probe; Lepr db/ + mice; genotype
                       
AuthorInstitution
赵英政 新乡医学院公共卫生学院,河南新乡
彭强 新乡医学院公共卫生学院,河南新乡
闫婷婷 新乡医学院公共卫生学院,河南新乡
张旭旭 新乡医学院公共卫生学院,河南新乡
翟晓楠 新乡医学院公共卫生学院,河南新乡
吴卫东 新乡医学院公共卫生学院,河南新乡
易宪文 新乡医学院公共卫生学院,河南新乡
徐光翠 新乡医学院公共卫生学院,河南新乡
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Abstract:
      Objective To establish an efficient method of genotyping for Lepr db/ + mouse offsprings by TaqMan probe quantitative fluorescence PCR. Methods Genome DNA was extracted from tails of 228 Lepr db/ + mouse offsprings. PCR primers and TaqMan probes were designed according to the mutation sites of Lepr gene (rs1801133). Real time PCR assay was applied and SNP loci were typed with SDS software. The genotyping of 2-month old Lepr db/ db mice was validated by the phenotype and Hardy-Weinberg equilibrium test was performed. Results 228 samples were detected by the established TaqMan fluorescence quantitative PCR assay. 64 mice were of GG genotype, with a genotype frequency of 0.1929. 123 mice were of GT genotype, with a genotype frequency of 0.5395. 41 mice were of TT genotype, with a genotype frequency of 0.2807. Compared with the phenotype typing, the sensitivity of the TaqMan fluorescence quantitative PCR was 97.56% and the specificity was 99.47%. Conclusions TaqMan probe quantitative fluorescence PCR assay is a simple and efficient method, and can be used to detect the genotype of Lepr db/ + mouse offsprings.
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