A method for specific detection of high-throughput transformants using KASP technology

A transformant, high-throughput technology, applied in the field of molecular biology, to achieve high accuracy, save manpower and time, and reduce the probability of errors

Active Publication Date: 2020-12-15
HUAZHI RICE BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, there is no report on the use of KASP technology for the specific detection of transformants in transgenic crops.

Method used

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  • A method for specific detection of high-throughput transformants using KASP technology
  • A method for specific detection of high-throughput transformants using KASP technology
  • A method for specific detection of high-throughput transformants using KASP technology

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Embodiment 1

[0055] Example 1 The method of the present invention is used to perform high-throughput detection on samples with known genotypes.

[0056] When the method of the invention is used for detection, the primer of the target sequence of the transformant and the primer of the endogenous gene are added into the reaction system, so how to ensure that two pairs of primers do not interfere with each other is a difficult point in this application. In addition, when the method of the invention is used for high-throughput detection of a large number of samples, the accuracy and reliability of the detection results is another difficulty. In order to test the effect of primers in the method of the invention and the accuracy of high-throughput detection by the method of the invention, transformant-specific detection is carried out on samples of corn, rice, soybean and rape with known genotypes, and then the detection results are compared with the genotype information of the samples themselves. T...

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Abstract

The invention provides a high-throughput transformant specificity assay method based on a KASP (Kompetitive Allele Specific PCR) technology. The assay method comprises the following steps: (1) primersof transformant target sequences and primers of endogenous genes are designed; (2) DNAs of samples are extracted; (3) PCR (polymerase chain reaction) is carried out by utilizing the primers of the transformant target sequences, the primers of the endogenous genes, KASP reaction solution and the DNAs of the samples to judge whether the samples contain the ingredients of transformants. When utilized to carry out transformant specificity assay on plant samples, the method has the advantages of high throughput, low cost, high accuracy, easiness in operation and the like.

Description

Technical field: [0001] The invention relates to the field of molecular biology, in particular to a method for high-throughput transformant specificity detection by using KASP technology. Background: [0002] Molecular detection of genetically modified crops and their products is the basis of safety management of genetically modified organisms. At present, China's imported genetically modified raw materials include dozens of transformants of corn, soybean, rape and other crops, and quite a few transformants can't be imported temporarily because they haven't been approved by the relevant state departments. Transformant-specific detection has at least three functions: First, it can prevent the import of unapproved transformants, which is beneficial for our country to effectively supervise the imported transgenic crops and their products; Secondly, the detection of transformant specificity is one of the means to detect the authenticity of transgenic varieties. Thirdly, if quantitati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6858
CPCC12Q1/6858C12Q2563/107C12Q2545/101C12Q2531/113
Inventor 杨翅春郑秀婷
Owner HUAZHI RICE BIO TECH CO LTD
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