摘要
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 develop a method for quantifying gene expressions in the mouse single oocyte and preimplantation embryo. Methods ： We quantified the message RNA （mRNA） expression of the TSC2 gene in the single oocyte and preimplantation embryo by capillary electrophoresis using the exogenic mutation TSC2 gene as the reference and amplification by competition polymerase chain reaction （PCR）. Results ： We successfully established the method for quantifying the mRNA expression of the TSC2 gene, with good linear relations between the mRNA level of the TSC2 gene and the dilution degree of the reference gene （ r = - 0. 987 ）. Conclusion： The level of the mouse TSC2 gene expression can be effectively quantified by competition PCR and capillary electrophoresis, which has provided a molecular base for evaluating the quality of human oocytes and preimplantation embryos.
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2009年第5期417-421,共5页
National Journal of Andrology
基金
南京市卫生局科研课题(YKK06101)~~
