实时荧光定量PCR检测传代培养细胞Grb10的表达

Detection of Grb10 Gene Expression in Cultured Cells by Real-Time PCR

目的建立一种可靠的Grb10印迹基因实时荧光定量PCR方法,检测传代培养鼠尾成纤维样细胞中Grb10表达的差异。方法通过实时荧光PCR,选择合适的"相对标准品"绘制相对标准曲线,检测传代培养细胞中印迹基因Grb10的表达水平。结果相对标准曲线有较好的线性(r=0.999及0.984);实时荧光定量PCR方法检测出随着细胞培养代数的增加印迹基因Grb10表达水平上升。结论双标准曲线的实时荧光定量PCR法用于检测Grb10的表达准确可靠;传代培养可能影响鼠尾成纤维样细胞中印迹基因Grb10的表达水平。

Objective Establishment of a reliable method of real-time PCR to detect the expression of imprinted gene GrblO in serial cultured fibroblast-like cells from mouse tail-tip. Methods Establishing the method of real-time PCR by selecting the suitable ＂relative standard preparation＂ and drawing the relative standard curve,which is used to detect expressive level of Grb10 in serial subcultivated fibroblast-like cells. Results The correlation coefficient of standard curve were 0. 999 and 0. 984 respectively. By the establishment of the method of real-time PCR,we detect that the expressive level of Grb10 rises in the process of serial subcultivation. Conclusions It is precise and dependable to detect the expression of GrblO by the method of real-time PCR of double standard curve. Serial subcultivation effect the expressive level of Grb10 in fibroblast-like cells from mouse tail-tip.