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本文针对转基因苜蓿草品系J101,设计引物及探针建立J101品系特异实时定量PCR检测体系。结果表明,本文建立的定量实时PCR检测方法特异性良好。实时荧光定量PCR检测方法的检测下限为10拷贝J101基因组DNA,定量检测下限为50拷贝的J101基因组DNA,本方法建立的标准曲线线性相关系数(R2)为0.993,扩增效率E为110%,重复性实验显示本方法的标准偏差(SD)及相对标准偏差(RSD)都在可接受范围内。本文建立的J101品系特异性定量PCR检测方法适于快速、准确、稳定地对转基因苜蓿草J101品系进行检测。
In this paper, the transgenic alfalfa grass J101, primers and probes were designed to establish J101 strain specific real-time quantitative PCR detection system. The results showed that the method of quantitative real-time PCR established in this paper has good specificity. The detection limit of real-time PCR was J101 genomic DNA of 10 copies and the lower limit of J101 genomic DNA was 50 copies. The linear correlation coefficient (R2) of standard curve established by this method was 0.993, the amplification efficiency E was 110% Repeatability experiments showed that the method of standard deviation (SD) and relative standard deviation (RSD) are within acceptable range. The J101 strain specific quantitative PCR detection method established in this paper is suitable for the rapid, accurate and stable detection of the transgenic alfalfa J101 strain.