TaqMan探针与SYBR Green实时定量PCR法检测转基因植物外源基因拷贝数的差异分析

Variance analysis between TaqMan probe and SYBR Green real-time quantitative PCR for determining the copy number of transgene in transformed plants

探讨TaqMan探针与SYBR Green实时定量PCR 2种方法检测转基因植物外源基因的拷贝数方法的差异。以12株T0转基因水稻为材料,分别用TaqMan与SYBR Green实时定量PCR法检测其拷贝数,然后用SAS 9.1软件对2种方法的结果进行t检验分析。通过SAS对2组数据的t检验分析,在内参基因扩增效率与目的基因扩增效率接近时,SYBR Green与TaqMan探针法的结果接近,差异不显著;当内参基因与目的基因扩增效率差异明显时,SYBR Green与TaqMan探针法差异显著。在内参基因扩增效率与目的基因扩增效率接近时,SYBR Green法测定转基因植物外源基因的拷贝数时结果接近TaqMan探针法。

To explore the differences between TaqMan probe and SYBR Green real-time quantitative PCR technique for estimating the transgene copy number in a transgenic plant, we used TaqMan probe and SYBR Green real-time quantitative PCR technique to determine the copy number of twelve transgenic flee, and then used the software SAS 9.1 to analyze the results with the two methods. As a result, the t-test analysis showed that when the amplification efficiency of the endogenous reference gene was close to the target gene, the variances between TaqMan probe and SYBR Green real-time quantitative PCR technique were not significant; As the amplification efficiency of the endogenous reference genes was in accord with the target gene, the results of the two methods were similar.