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Development of Digital PCR Absolute Quantification Method for Herbicide-tolerant Genetically Modified Soybean SHZD32-1 |
ZHENG Zi-fan1,WANG Hao-qian2,GAO Jia-qi3,CHEN Shuo3,LIU Fang-fang3,ZHANG Xiu-jie2,LI Liang1,3 |
1. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
2. Development Center of Science and Technology, Ministry of Agriculture and Rural Affairs P. R. China, Beijing 100025, China
3. Institute of Quality Standard and Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing 100081, China |
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Abstract Based on the digital PCR technology for absolute quantification of DNA single molecule, a droplet digital PCR (ddPCR) dual (transformant-specific gene and soybean internal standard gene) quantitative method for the transformant was established. The main contents include:optimize the double ddPCR primer-probe concentration and annealing temperature, examine the specificity, determine the linear kinetic range and critical parameters such as the limit of detection (3copies/μL) and the limit of quantification (15copies/μL). The genetically modified soybean SHZD32-1 has obtained the production and application safety certificate "Nongjianzhengzi (2019)" No. 293. Its ddPCR quantitative detection method, as an absolute quantitative measurement method, is simple, efficient and accurate, and will provide technical support for the calibration of reference materials, quantitative detection and standardization of the establishment of digital PCR methods in the field of biosafety.
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Received: 21 February 2022
Published: 08 March 2023
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