Method for Quantitative Determination of Genetically Modified Sugar Beet Line H7-1 by Using Digital Polymerase Chain Reaction
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Abstract:
In order to achieve labeling management and accurate quantification of genetically modified sugar beet H7-1, primer pairs and probes based on the 5? flanking sequence and glutamine synthetase (GS) of H7-1 were designed, and a duplex digital polymerase chain reaction (dPCR) detection method for H7-1 was established. The specificity, sensitivity, precision, and accuracy of the developed method were examined. The results showed that the developed dPCR method was specific for line H7-1 detection. The limits of quantitation (LOQs) of the specific sequence and endogenous GS gene of line H7-1 in a 20-μL reaction system were 3.1 copies/μL and 6.3 copies/μL respectively. The limits of detection (LODs) of the specific sequence and endogenous GS gene of line H7-1 in a 20 μL reaction system were 0.6 copies/μL and 1.3 copies/μL respectively. The precision and accuracy were all in the acceptable range. This quantitative method does not rely on the establishment of a standard curve, and can be easily applied for accurate quantification of sugar beet H7-1.
