Quantitative Analysis of Genetically Modified Rapeseed of RF1 by Duplex Droplet Digital Polymerase Chain Reaction (Duplex-ddPCR)
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Abstract:
A duplex droplet digital PCR (duplex-ddPCR) quantitative analysis method for the the simultaneous detection of genetically modified rapeseed Rf1 was established based on the droplet digital PCR platform. The results showed that both endogenous reference gene and Rf1 event-specific gene could be specifically amplified, and the established duplex-ddPCR method for RF1 event-specific rapeseed had good specificity. In unit system, the reference gene number and the exogenous gene copy number showed a good linearity (r2=0.999) in the range of 18~23077 copies, andthe LOQ and LOD were 18 copies and 3.7 copies, respectively. The precision test results showed that the relative standard deviations (RSD) of endogenous PEP and RF1 content were between 8.40% and 24.50%, respectively, and the accuracy test results showed that the relative standard deviations (RSD) were between 5.97% and 12.64%, respectively. The RSD of precision and accuracy were all less than 25%, which met the detecting requirements. In conclusion, the duplex-ddPCR quantitative analysis method established in this study could be used for the quantitative detection of genetically modified rapeseed RF1.
