Lamp detection primer set, lamp detection kit and detection method of transgenic insect-resistant soybean mon87701 and its derivatives

Abstract

The invention discloses a LAMP detection primer set, a LAMP detection kit and a detection method of the transgenic insect-resistant soybean MON87701 and its derivative varieties. The LAMP detection primer set includes four specific primers; the detection kit includes a primer solution, a reaction solution, and a DNA polymerase. and control; the kit can also have a chromogenic agent or a fluorescent indicator; the detection method is to extract the DNA of the sample to be tested, using four specific primers and a DNA polymerase with strand displacement activity, at 60-65 ° C Amplify the sample DNA template, and the short-term amplification efficiency can reach 10 9 ~10 10 The identification uses the turbidity change of the precipitate in the reaction tube, adding a chromogen to observe the color change in the reaction tube, and using a real-time fluorescence detector to observe and determine whether the sample to be tested contains transgenic insect-resistant soybean MON87701 and its derivatives. The invention has the advantages of rapidity and high efficiency, simple operation, high specificity, high sensitivity, simple identification, suitable for on-site detection and the like, and is suitable for popularization and application.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a LAMP detection primer set, a LAMP detection kit and a detection method of transgenic insect-resistant soybean MON87701 and its derivative varieties. Background technique [0002] Transgenic technology was first successfully developed in 1973 by Tanley Cohen, a professor at Stanford University. Genetically modified soybeans are one of the examples of the successful application of genetically modified technology in crops. The world's first genetically modified soybean plant was born in 1988. In 1994, the glyphosate-resistant genetically modified soybeans launched by Monsanto Company of the United States were approved for commercial production. Soybean planting area expanded rapidly. The application of transgenic technology and the commercialization of global transgenic plant cultivation provide an effective way to solve the problem of food shortage. [0003] Soybean i...

AI Technical Summary

Problems solved by technology

[0007] 1) Protein-based detection, now the most widely used detection methods are enzyme-linked immunosorbent assay (ELISA) and Western blot detection methods, which require high-quality, high-specificity antibodies, otherwise it will affect the accuracy of detection, low sensitivity;

[0008] 2) DNA-based detection is mainly for the detection of exogenously inserted genes in transgenic crops, mainly including molecular hybridization technology, PCR detection technology and chip detection technology, among which PCR detection method is the most important and most accurate detection of genetically modified crops method, but the reaction system and operation process of this type of method are relatively complicated, and professionals are required; a specific PCR instrument is required, and the amplification time is 2 to 3 hours. Strong toxicity, and difficult to implement on-site detection

[0010] Loop-Mediated Isothermal Amplification technology (Loop-Mediated Isothermal Amplification, hereinafter referred to as LAMP method) is a gene amplification technology developed by Notomi and Okayama around 2000. It has the advantages of fast and simple, accurate operation, easy popularization, safety and reliability. At present, there is no kit that applies constant temperature amplification visual detection technology and constant temperature real-time fluorescence technology to the rapid detection of transgenic insect-resistant soybean MON87701 and its derivatives

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