Research On Event-specific Analysis Of Genetically Modified Rapeseed Detection And Development Of Reference Molecules

In order to analyze the composition of genetically modified (GM) rapeseed events in Shanghai port,27 lots of rapeseed samples imported from Canada and Ukraine were analyzed. Event-specific real-time PCR methods suitable for GM rapeseeds RT73, T45, Oxy235, Topas19/2, MS1×RF1, MS1×RF2 and MS8×RF3 detection were employed. Results showed that seven GM rapeseed events certificated in import by Ministry of Agriculture of People's Republic of China have all been imported from Shanghai port. GM rapeseed events RT73 and MS8×RF3 were the major ones.Due to the deficiency of reference materials for GMO analyses, reference molecules pEASY-RT73 and pCanola9 suitable for event-specific analysis of GM rapeseed was developed. In identification and quantification of the reference molecule was also validated in our lab. Results showed that both of them were highly specific to rapeseed detection. In the qualitative PCR analysis using reference molecules as the calibrator, limit of detection (LOD) are all lower than 20 copies. In the real-time PCR, LODs for targets were all lower than 25 copies, and LOQs were all lower than 50 copies. Against the standard curve produced by the above system has a good linear relationshipand high reaction efficiency.Furthermore, research has been made on repeatability and reproducibility detection, thermostabilization and blind samples detection in reference molecules. The developed reference molecule pEASY-RT73 and pCanola9 could be used to qualitatively and quantita-tively analyze GM rapeseed and its derived products as a good alternative of reference material derived from plant materials.