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目的:建立在小鼠单个卵母细胞和早期胚胎中结节性硬化综合征2型(TSC2)基因mRNA定量检测的方法。方法:选择外源性内参,采用PCR技术,行微流电泳定量检测单个卵母细胞和早期胚胎中TSC2基因mRNA表达水平。结果:成功建立小鼠单个卵母细胞和早期胚胎中TSC2基因mRNA表达的定量检测方法,并绘制定量标准曲线,相关性好(r=-0.987)。结论:采用PCR技术,行微流电泳定量检测单个小鼠卵母细胞和早期胚胎中TSC2基因mRNA表达的方法可行,为从分子水平评价人类配子与胚胎质量提供了方法学基础。
Objective: To establish a method for the quantitative detection of TSC2 mRNA in mouse single oocytes and early embryos. Methods: Exogenous internal reference was selected and the expression of TSC2 mRNA in single oocyte and early embryo was determined by PCR and microfluidic electrophoresis. Results: Quantitative detection of TSC2 mRNA expression in single oocyte and early embryo of mice was successfully established and a quantitative standard curve was drawn. The correlation was good (r = -0.987). Conclusion: It is feasible to use PCR to detect the mRNA expression of TSC2 gene in single mouse oocytes and early embryos by using microfluidic electrophoresis, which provides a methodological basis for evaluating the quality of human gametes and embryos at the molecular level.