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本文以我国批准商业化的转基因耐草甘膦棉花MON88913为研究对象,建立并验证了其转化体特异性定性、定量PCR检测方法。建立的定性PCR方法的检测极限是20个拷贝棉花单倍体基因组DNA,定量PCR方法的检测和定量极限分别是10和20个拷贝棉花单倍体基因组DNA。同时,我们组织了实验室5位研究人员对建立的定量PCR检测方法进行了协同验证。对5个盲样的定量分析结果显示与真实值的偏差介于1.59%和10.12%之间,完全满足国际标准25%偏差范围的要求,完全可用于转基因棉花MON88913的实际样品检测。
In this paper, commercialized transgenic glyphosate-resistant cotton MON88913 was selected as the research object, and its transformant-specific qualitative and quantitative PCR detection methods were established and validated. The detection limit of the established qualitative PCR method was 20 copies of cotton haploid genomic DNA. The detection and quantitation limits of the quantitative PCR method were 10 and 20 copies of cotton haploid genomic DNA. At the same time, we organized a laboratory of five researchers to establish a quantitative PCR detection method for collaborative verification. Quantitative analysis of five blind samples showed that the deviation from the true value was between 1.59% and 10.12%, which completely met the requirement of 25% deviation from international standard and could be used for the actual sample detection of transgenic cotton MON88913.