Primer, reagent box and method for detecting transgenic soybean MON87708

Abstract

The invention discloses primer, a reagent box and a method for detecting transgenic soybean MON87708. The primer comprises LectinF: TCTGTCGTTGTGGTGCTA, LectinR: CTGCGAGATTTGCTATGT, MON87708F: GTCAAGGAGGACAAGGTCG and MON87708R: CCAATGCCATAATACTCAAAC. The reagent box is filled with 10 uL of HRM reaction premix liquid, 0.2-3.4 uL of primer, 1 uL of DNA template and is filled into 20 uL by water. The detecting method includes extracting sample DNA, and subjecting sample DNA in the step A to PCR (polymerase chain reaction) amplification by means of the reagent box; after amplification, products are subjected to high-resolution melting curve analysis, and results are judged by an amplification curve. In the primer, the reagent box and the method for detecting transgenic soybean MON877088 and endogenous genes Lectin, the reagent box is reasonable in component and proportion, is convenient to use and quick and accurate to detect, the method is convenient and quick to operate, and low in cost, and detecting results are accurate.

Description

technical field [0001] The invention designs a double fluorescent PC detection kit for detecting transgenic soybean MON87708 and endogenous gene Lectin, and the invention also relates to a method using the above kit for transgenic soybean MON87708 and endogenous gene Lectin. Background technique [0002] In the 1980s, researchers from the herbicide production company Monsanto cloned the resistance gene-EPSPS gene from petunia, which is the 5-enonepyruvate shikimate-3-phosphatase gene of Pseudomonas. Using Ti plasmid-mediated transfer DNA (TV-DNA technology), the EPSPS gene controlled by the 35S promoter in the petunia Ti plasmid CaMv was introduced into the soybean genome, and then the transgenic soybean variety Roundup Ready Soybean, which is resistant to the herbicide glyphosate, was bred. RR soybeans. This genetically modified soybean was approved by the U.S. FDA in 1994 and became one of the earliest genetically modified crops to be commercially promoted and produced on...

AI Technical Summary

Problems solved by technology

[0007] However, the conventional PCR method needs electrophoresis after PCR amplification, which is not only cumbersome to operate, but also easily causes pollution, and also needs to use fluorescent fuels that may cause cancer. Although the probe-based fluorescent PCR method is sensitive and accurate, the linear structure of the TaqMan probe leads to High background fluorescence, if the distance between the fluorophore and the quencher group is too close, the possibility of the probe degrading between them during the PCR amplification process will be greatly reduced, so that it will not function as a probe

On the contrary, if the fluorescent group and the quencher group are placed at both ends of the probe, the fluorescent background signal will be strengthened, which will affect the sensitivity of its detection.

The high specificity of TaqMan will lead to a base that also limits its detection target to be too narrow. In addition, its synthesis cost is high, and the reagent has a short validity period and is easy to degrade. Therefore, it cannot be widely used at the grassroots level.

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