关键词: 双重数字PCR, 转基因大豆, 定量检测

Abstract: In this study, a duplex PCR detection method for three genetically modified soybean of MON87705, MON87769 and DP356043 was established by droplet digital PCR (ddPCR) platform. Taking  advantage of specificity and quantitative range of duplex digital PCR, the primer probe combination and experimental procedure was optimized, exogenous genes and reference gene were detected. The experiment result showed that the primer probe combination used in the experiment only produced fluorescent signal for the target soybean line in the digital PCR method. It can be used in the detection of genetically modified soybean for its specificity. The genetically modified ingredients content was basically the same as reference parameters. The quantitative detection limit was 0.5%, and the qualitative detection limit was 0.05%, which could meet the needs of the detection of low-purity sample. The study also determined the gene copy number of transgenic soybean, which indicated that the dual digital PCR system established in this study can accurately and stably meet the actual detection needs, and has a good development prospect in practical application.

Key words: duplex droplet digital PCR, genetically modified soybean, quantitative detection