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Detection of Genetically Modified Soybean in Processed Foods Using Real-Time Quantitative PCR with SYBR Green I Dye on the DNA Engine Opticon 2 System

ak is present only in the spiked samples (Data not shown and Table 5) ruling out a false-negative result. It should be noted that the soy flour samples that are negative for the EPSPS gene might still contain other types of GMOs that were not being tested in this assay.

To confirm that the GMO-positive food samples did contain the EPSPS gene, the amplified PCR products from reactions with the EPSPS primers were sequenced. As expected, a BLAST search with the obtained DNA sequence showed 100% concordance with the sequence of the glyphosate tolerant version of the cp4 epsps gene from Agrobacterium tumifaciens (GenBank Accession AF464188). The reaction products were also examined on an agarose gel. The band pattern for all reactions was in complete agreement with the results of the real-time qPCR data (not shown).


Discussion
In this study, we have developed a real-time qPCR protocol for the DNA Engine Opticon 2 system that can be used to detect and quantify genetically modified soy. The GMO detection assay presented here has several features that make it attractive for rapid processing of multiple samples. The amount of required starting material is quite small. We have found that between 50 to 100mg of a given food sample yields more than enough DNA for testing. Detection with SGI can be used for any DNA amplicon, allowing this protocol to be easily adapted for detection of GMOs in other crops such as maize, peanut and rice.


References
1. Schonbrunn, E., Eschenburg, S., Shuttleworth, W.A., Schloss, J.V., Amrhein, N., Evans, J.N.S. and Kabsch, W. Interaction of the herbicide glyphosate with its target enzyme 5-enolpyruvyls
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