Method, reagent and kit for detecting transgenic maize line 98140

Abstract

The invention discloses a method, reagent and kit for detecting a transgenic maize line 98140. The detection method comprises the steps that specific primers and probes of a line 98140 exogenous geneand HMG gene are simultaneously added into a PCR reaction system, mineral oil is added, uniform mixing is carried out to prepare micro-droplets, dual ddPCR is performed in the micro-droplets, after the reaction is completed, each micro-droplet amplification signal is read, and copy numbers of the line 98140 exogenous gene and HMG gene are calculated, wherein the probes of the maize line 98140 andHMG gene are fluorescently marked by FAM and HEX respectively. The method, reagent and kit for detecting the transgenic maize line 98140 have the advantages that the endogenous gene HMG and line specific primer probes which are matched for use adopt a two-channel method to quantify the copy numbers of the line 98140 exogenous gene and HMG gene in a maize genome separately for calculating the absolute content of the transgenic maize line 98140, thereby having great significance to studying the transgenic components of maize and accurately and quantitatively detecting the transgenic components in imported and exported maize products.

Description

technical field [0001] This application relates to the field of transgenic maize detection, in particular to a method, reagent and kit for detection of transgenic maize line 98140. Background technique [0002] With the rapid development of the biotechnology industry, the research and development speed and planting area of ​​genetically modified crops continue to increase globally. Promoting genetically modified technology has become an inevitable choice for future-oriented development strategies and to solve food shortages and ensure national food security. However, the safety of genetically modified organisms has also become a hot spot in the world. At present, all countries in the world have strengthened the management and testing of genetically modified organisms and their products for various purposes such as economy, health, and environmental protection. About 50 countries and regions, including China, have promulgated and implemented the "GMO labeling system" to lab...

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Problems solved by technology

However, digital PCR is still in its infancy in the research of genetically modified detection, especially in the standard level of my country and even countries around the world, which greatly restricts the application of digital PCR in actual detection.

[0005] Studies have shown that, according to the statistics of the planting area, about 35% of the world's corn is genetically modified; however, international and domestic specific qualitative or quantitative detection methods for genetically modified corn are quite lacking, especially for the quantitative detection of the genetically modified corn line 98140. There are relevant studies or reports

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