转基因苜蓿草KK179-2品系特异性实时荧光PCR检测方法的建立

发布时间：2018-06-22 23:32

  本文选题：转基因苜蓿KK- + 品系特异性　； 参考：《中国饲料》2017年14期

【摘要】：根据转基因低木质素苜蓿草品系KK179-2 5’端外源插入序列与苜蓿草基因组DNA之间的邻接区序列设计引物和探针,建立了KK179-2品系特异性实时荧光PCR检测方法,并对其特异性、灵敏度及可重复性进行了测定。结果表明:建立的定量实时荧光PCR检测方法特异性良好;标准曲线线性相关系数(R~2)为0.99,扩增效率E为105%;检测限为20拷贝,定量限为200拷贝。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。综上,建立的KK179-2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KK179-2品系进行检测。
[Abstract]:The primers and probes were designed according to the sequence of the adjacent region between the exogenous insertion sequence at the end of KK179-2 5'terminal and the genomic DNA of alfalfa genome of transgenic alfalfa line KK179-2, and the specific real-time fluorescent PCR detection method for KK179-2 strain was established and its specificity was analyzed. Sensitivity and repeatability were determined. The results showed that the specificity of the method was good, the linear correlation coefficient of standard curve was 0.99, the amplification efficiency E was 105, the detection limit was 20 copies and the quantitative limit was 200 copies. Repeatability experiments showed that the standard deviation (SD) and relative standard deviation (RSD) of the method were within acceptable range. In conclusion, the established KK179-2 strain specific quantitative PCR method is suitable for rapid, accurate and stable detection of transgenic alfalfa line KK179-2.
【作者单位】： 黄埔出入境检验检疫局;广东出入境检验检疫局;
【基金】：广东出入境检验检疫局科技项目(2017GDK41、2016GDK60)
【分类号】：S541.9

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