A New Approach For Rapid Extraction And Fast Detection Of Genetically Modified Organisms

As more and more GM crops were approved for commercialization and planting, the development of quick and on-spot analysis methods for GM crops and their derivates are very important and useful. Loop mediated isothermal amplification (LAMP) is one isothermal nucleic acids amplification technique. The LAMP reaction shows the higher specificity, simplicity, rapidness, compared with typical PCR. It is no need of the expensive PCR equipments and argarose gel electrophoresis (AGE) assays. The technique is less-consuming time for DNA amplification (within 60 min) and high efficiency by direct visual inspection, which can be used for large numbers of GMOs detection on spot. Furthermore, the existing extraction DNA methods for transgenic crops are consuming time(about 90 min). Simultaneously, it is required for the unwieldy centrifuge instrument taking the extraction. We studied a novel technology for extracting of DNA to reduce the time from the existing about 90 minutes to about 10-15 minutes. Combined the fast extracting DNA technology with the LAMP method, the extraction and detection can be completed within 90 minutes for on spot detection of GMOs.1,Based on the event-specific sequences of seven GM maizes (DAS-59122-7, T25, BT176, TC1507, MON810, BT11 and MON863), LAMP event-specific primers were designed using the specific software of Primer Explorer V4. In this study, the optimized visual LAMP methods for the detection of GM maize events have been established. In combination with SYBR Green I dye, the amplified products can be directly detected by naked eye instead of the conventional gel electropherisis anlaysis. The limits of detection (LODs) were as low as four copies of maize haploid genomic DNA except for the forty copies of MON810 assay. Furthermore, these developed assays were successfully used in the USDA/GIPSA Proficiency Program, all the six mixed maize samples with different contents (ranged from 0.0% to 2.0%) of GM maize events were identified correctly. The LAMP assay has the advantages of time-saving and low cost or simple procedures for result analysis, and can be used for GMOs detection on spot.2,Research on the rapid extraction of DNA for GM organism. We designed simple equipment for extracting DNA, which contains a simple injector instead of the unwieldy centrifuge instrument. The technology used principle of atmospheric pressure, instead of weightlessness. We studied the novel technology for extracting DNA to reduce the time from the existing about 90 minutes to about 10-15 minutes. The technology was employed to extract the rice, the maize, the cotton and the soybean, and was used to fast extraction on spot. Combined the fast extracting DNA technology with the LAMP method, the method can be used for GMOs detection on spot.