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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

on of glyphosate to fields of the growing GMO crop, e.g., Roundup Ready soybeans.

Because GMOs contain specific and well-defined nucleic acid sequences, they are well suited for detection by the polymerase chain reaction (PCR). Indeed, PCR has been shown to effectively detect GMO material, using an agarose gel analysis of the reaction products.4, 5

In this Application Note, we extend the PCR detection of GMOs by presenting a real-time qPCR protocol using SYBR Green I (SGI) performed on the DNA Engine Opticon 2 continuous fluorescence detection system. DNA was isolated from reference materials containing known proportions of GMO soy, as well as from soy products obtained from a local supermarket. Control primers that amplified a portion of the soy lectin gene were used to normalize the amount of soy DNA in each sample. A second set of primers that amplify a portion of the glyphosate-tolerant EPSPS gene were used for GMO detection.

The threshold cycle [C(t)] of a reaction is defined as the point when a reaction signal just rises over the background signal level. Comparing the relative C(t)s of lectin and EPSPS reactions from the GMO concentration reference series allowed the construction of a standard curve, from which the amount of GMO materials in unknown foodstuffs could be determined down to approximately 0.5% (w/w) concentration.


Materials and Methods
Isolation of DNA from Reference Standards and Food Samples
Dried soybean powder containing mass fractions of 0% to 5% Roundup Ready soybean (RTC Corporation, IRMM-41050-56) were used as reference standards. The reference standards were prepared by the manufacturer by mixing dried non-genetically modified and
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