中国实验动物学报2018，Vol.26Issue(2)：207-210,4.DOI:10.3969/j.issn.1005-4847.2018.02.011
应用TaqMan探针荧光定量PCR技术鉴定Leprdb/+小鼠子代基因型
Genotyping of the offsprings of Leprdb/ +mice by TaqMan probe fluorescence quantitative PCR
摘要
Abstract
Objective To establish an efficient method of genotyping for Leprdb/ +mouse offsprings by TaqMan probe quantitative fluorescence PCR. Methods Genome DNA was extracted from tails of 228 Leprdb/ +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 Leprdb/dbmice 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 Leprdb/ +mouse offsprings.关键词
Taqman探针/Leprdb/+小鼠/基因分型Key words
TaqMan probe/Leprdb/ +mice/genotype分类
生物科学引用本文复制引用
赵英政,彭强,闫婷婷,张旭旭,翟晓楠,吴卫东,易宪文,徐光翠..应用TaqMan探针荧光定量PCR技术鉴定Leprdb/+小鼠子代基因型[J].中国实验动物学报,2018,26(2):207-210,4.基金项目
国家自然科学基金面上项目(No.81370916,No.81773399),新乡医学院科研培育基金(No.2014QN110).Funded by National Natural Science Foundation of China(No.81370916,No.81773399),Scientific Research Cultivation Fund of Xinxiang Medical University(No.2014QN110). （No.81370916,No.81773399）
