【摘要】 根据GenBank中卤虫actin基因的保守区域序列设计并合成一对引物,采用SYBR GreenⅠ作为染料建立了实时荧光定量PCR(real-time fluorescence quantitative PCR)方法.以Ct值为纵坐标,以稀释倍数的对数为横坐标,建立标准曲线,其相关系数达到了0.999.溶解曲线分析显示产物为单一的特异峰,Tm为83.6℃.结果表明:本实验建立的中国卤虫actin基因的实时荧光定量PCR法扩增效率高、特异性强、线性范围广、检测周期短,为actin基因作为内参基因进行卤虫早期胚胎发育基因表达的定量分析奠定了基础.更多还原

【Abstract】 According to the actin gene sequences of Artemia sinica available in GeneBank,a pair of primers was designed for establishing a SYBR Green I quantitative real-time fluorescence quantitative PCR method.Standard curve was established basing on Ct vs initial input amount of Log(copy number),and the correlation coefficient of it was 0.999.The melting curve showed a single peak with a T_m of 83.6 ℃.The results demonstrated that method developed in this study can quickly detect actin in broad range with high efficiency,specificity.The experiment provided the basis for actin gene of Artemia sinica as a control gene in quantitative analysis of early embryonic development gene expression of Artemia sinica.更多还原