Establishment of Precisely Quantitative Method of Genetically Modified Rice LL62 Based on Digital PCR

doi:10.13560/j.cnki.biotech.bull.1985.2016.08.011

Abstract

Abstract: In order to promote the smooth implementation of labeling policy for genetically modified（GM）products,we study an precise quantitative detection method for the GM rice LL62 that has not been approved by the Ministry of Agriculture in China,aiming at guarantee the detection and supervision of GM agricultural products in export and import. A digital PCR（dPCR）detection system were designed based on the inserted exogenous fragment of LL62 and 3’ flanking sequence of DNA of rice genome,and a precisely quantitative method was established. The results showed that the designed probes presented high efficiency,the developed dPCR method was highly specific for GM rice LL62 detection. The method was highly repeated,and the relative standard deviation（RSD）values of droplets’ number ranged from 0.60%-11.11%. Results of the quantification of three blind samples showed that the bias between the true value and the measured value was 0.12,0.09 and 0.10,the RSD was 0.09%-10.31%,indicating that the accuracy was high. In conclusion,the established droplet dPCR method for GM rice LL62 detection is simple and convenient with high specificity,solid repeatability and high accuracy,and is suitable for precisely and effectively quantitative analysis of ingredients of GM rice or peeled rice LL62 in agricultural products and foods.

Key words: genetically modified rice LL62, droplet digital PCR, event-specific, quantification

REN Yi-fei, GAO Qin, DENG Ting-ting, LI Xiang, HUANG Wen-sheng, CHEN Shun-sheng, CHEN Ying. Establishment of Precisely Quantitative Method of Genetically Modified Rice LL62 Based on Digital PCR[J]. Biotechnology Bulletin, 2016, 32(8): 69-76.

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