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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

roduct with the EPSPS primers, the ΔC(t) value was designated as not detected. Next, a standard curve was generated by plotting the log %GMO concentration of the reference standards against the ΔC(t) value (see Figure 1B). In order to calculate the %GMO content of an unknown food sample from the standard curve, the Excel Trend function was used. In the Trend function, the following values were specified for the standards: log %GMO (the known y values), ΔC(t) (the known xs) and the ΔC(t) of the unknown food sample for the new x. The value being generated from the formula is the log %GMO of the unknown. The %GMO soy content of the unknown sample was then determined from the formula 10 (log%GMO).

Thermal denaturation profiles of all reactions were examined to verify the presence of the lectin amplicon (in all soy) and the EPSPS amplicon (GMO soy specific).


Results
A standard curve generated from the soy reference standards and the EPSPS primers is shown in Figure 1A. The linear range of the curve extended from 5% mass fraction of GMO soy standard down to 0.5% mass fraction of GMO soy standard, with the square of the correlation coefficient (r2) equal to 0.992. The average C(t) values for duplicate reactions of each sample DNA with the lectin primers and the EPSPS primers are listed in Table 3. The amount of GMO soy was normalized to the amount of total soy present by calculating the ΔC(t) value as described in the Materials and Methods section.

The standard curve generated by plotting the log %GMO of the standards against the ΔC(t) values of the standards is shown in Figure 1B. The %GMO soy
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