标准编号：ISO/TS 21569-4:2016
中文名称：分子生物标志物分析的水平方法 检测转基因生物体和衍生产品的分析方法 第4部分：P-nos和P-nos-nptII DNA序列检测用实时PCR筛选法
英文名称：Horizontal methods for molecular biomarker analysis — Methods of analysis for the detection of genetically modified organisms and derived products — Part 4: Real-time PCR based screening methods for t
发布日期：2016-11
标准范围
This document specifies a procedure for the detection of a DNA sequence of the promoter region of thenopaline synthase gene (P-nos) from Agrobacterium tumefaciens and a procedure for the detection of theDNA transition sequence between P-nos and the neomycin-phosphotransferase gene (nptII) from theTn5 transposon of Escherichia coli K12. The nos-promoter and the P-nos-nptII-construct are frequentlyfound in genetically modified plants. The P-nos and P-nos-nptII specific methods are based on realtimePCR and can be used for qualitative screening purposes. For identification and quantification of aspecific genetically modified plant (event) a follow-up analysis has to be carried out.The methods described are applicable for the analysis of DNA extracted from foodstuffs. They may alsobe suitable for the analysis of DNA extracted from other products such as feedstuffs and seeds. Theapplication of these methods requires the extraction of an adequate amount of amplifiable DNA fromthe relevant matrix.The DNA sequence amplified by the P-nos element-specific method can be detected in samples whichcontain DNA of the naturally occurring Ti-plasmid of A. tumefaciens. For this reason, it is necessary toconfirm a positive screening result. Further analyses are required using construct-specific or eventspecific methods.
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