为建立膜连蛋白 A2(AnxA2)基因 SYBR Green Ⅰ实时荧光定量 PCR 检测方法,根据 Gen-Bank 中公布的 AnxA2基因序列设计并合成引物,经过条件优化,建立了 AnxA2 SYBR Green Ⅰ实时荧光定量 PCR 检测方法。该方法线性关系较好,以质粒标准品构建的标准曲线相关系数 R2为0.999;灵敏性好,比普通 PCR 灵敏100倍。应用建立的方法对随机选取的几种哺乳动物细胞进行检测。结果表明,建立的方法能成功检出不同细胞中 AnxA2含量。表明本研究建立的 AnxA2 SYBR Green Ⅰ实时荧光定量PCR 检测方法灵敏性高,重复性好,可用于 AnxA2的检测及定量分析。
In order to establish AnxA2 SYBR Green I real-time PCR,one pair of primers were designed and synthesized based on the published gene sequence of AnxA2 in GenBank.After optimizing the reaction con-ditions,AnxA2 SYBR Green I real-time PCR was established with good linear relationship,and the stand-ard calibration curve correlation coefficient R2 was 0 .999 for standard plasmid.It also had good sensitivity, about 100 times more sensitive than conventional PCR.We also detected several randomly selected mam-malian cells with this method.The results showed that the method could successfully detect the content of AnxA2 in different cells.It proved that the established AnxA2 SYBR Green I real-time PCR was of high sensitivity,good reproducibility and could be used in the detection of AnxA2 and quantitative analysis.