Method for detecting paddy rice transgenic ingredients

Abstract

The invention discloses a method for detecting paddy rice transgenic ingredients, and belongs to the field of biotechnology. The method comprises: determining a transgenic ingredient, an endogenous standard gene and an exogenous standard gene; preparing a multiple-amplification primer; carrying out sampling and mixing to obtain a mixed sample; extracting a genome of the mixed sample, and adding the exogenous standard gene to obtain mixed nucleic acid; constructing a high-flux sequencing library; carrying out high-flux sequencing on the high-flux sequencing library to obtain a sequencing fragment group; obtaining the number of sequencing fragments of the transgenic ingredient, the number of sequencing fragments of the endogenous standard gene and the number of sequencing fragments of the exogenous standard gene in a to-be-tested sample; according to the number of the sequencing fragments of the exogenous standard gene, determining whether an experiment is successful or not, and calculating the content of the transgenic ingredient in the to-be-tested sample; and according to the content of the transgenic ingredient, determining whether the to-be-tested sample contains the transgenic ingredient. According to the method, various transgenic ingredients in the to-be-tested sample can be quantitatively detected at the same time.

Description

Technical field [0001] The invention relates to the field of biological detection, in particular to a detection method of rice transgenic components. Background technique [0002] Rice is the main food crop in my country, and the genetically modified technology of rice and its products has matured. The planting and promotion of genetically modified rice varieties require strict approval. Therefore, genetically modified rice needs to be supervised, and genetically modified component testing is the technical support for genetically modified supervision. [0003] The existing genetically modified component detection technology mainly detects nucleic acid or protein. Among them, the main process of nucleic acid-based detection methods is: extract nucleic acid from the sample to be tested, and use PCR (Polymerase Chain Reaction, polymerase chain reaction) method to amplify the transgene in the sample Ingredients, using real-time quantification, agarose electrophoresis or chip technology...

AI Technical Summary

Problems solved by technology

[0005] There are many kinds of genetically modified ingredients, and the existing technology can only detect one or a few of them at a time. Therefore, it is necessary to test the possible multiple genetically modified ingredients one by one before they can be determined as "non-genetically modified" products; The transformed genetically modified components are not the exogenous gene itself. Therefore, for unmarked transgenes, the existing detection methods will be invalid; detection is basically qualitative detection, but quantitative detection has its practical needs, for example, A small amount of pollen from genetically modified rice varieties in adjacent fields has been transferred to non-transgenic varieties. During qualitative testing, non-transgenic varieties will be misjudged as genetically modified varieties and will not be approved or given administrative penalties.

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