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| Establishment of a Novel Event-specific Quantitative PCR Method for Genetically Modified Cotton (Gossypium sp.) GHB119 Detection |
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Abstract To establish the detection method of genetically modified (GM) cotton event GHB119, which has not been authorized by the Ministry of Agriculture of China, the specific primer pair and probe were designed based on the 3'flanking sequence between the cotton genome and inserted exogenous fragment of GHB119 for protecting and improving the implementation of laws and regulations for GM products safety. After optimizing the reaction conditions, a novel event-specific quantitative Real-time PCR method for GHB119 detection was established. The applicability of the method was determined. Results showed that the developed Real-time PCR method was specific for GM cotton GHB119 detection, the limit of detection (LOD) and quantification (LOQ) were 10 and 25 copies of GHB119 haploid genomic DNA, respectively. Results of the quantification test of GM ingredient in blind samples showed that the bias between the true value and the measured value, the standard deviation (SD) and the relative standard deviation (RSD) were all in the acceptable range. Therefore, the established event-specific quantitative Real-time PCR method for GM cotton GHB119 detection has high specificity and good sensitivity, and is suitable for quantification of GHB119 ingredient in mixed samples quickly and accurately.
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Received: 13 September 2013
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