Dual PCR experiment

Summary

With the large-scale cultivation of genetically modified (GM) crops, the safety of GM crops has received increasing attention, and in 2002, China required the labeling of GM crops. Glyphosate-resistant soybean (trade name, roundup ready soybean) was obtained by transferring the exogenous gene CAMV-35S promoter, glyphosate-resistant gene CP4-EPSPS and NOS terminator into the traditional soybean strain Variety A5403. It is one of the most widely used transgenic crops, and at least 8 million tons of transgenic soybeans enter our market every year. Therefore, it is of great significance to study the rapid detection method of glyphosate-resistant soybean. Most of the current methods for detecting transgenic soybeans by PCR use agarose gel electrophoresis, which first detects the CAMV-35S promoter and NOS terminator for screening, and then detects the EPSPS glyphosate-resistant gene, which is a cumbersome and time-consuming operation. In this study, we utilized dual PCR to amplify the above gene fragments simultaneously, and then detected the PCR products by laser-induced fluorescence-capillary electrophoresis in an efficient and rapid manner, which significantly improved the detection efficiency and greatly increased the sensitivity. The method is rapid, sensitive and accurate, and has been successfully applied to the analysis of actual transgenic soybean samples with low sample volume (nl level).

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