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A dna sequencing method for a specific region of the genome

A DNA sequencing, specific region technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of inability to accurately obtain genotype DNA sequence information, and achieve simple design, high measurement throughput, and experimental design. simple effect

Active Publication Date: 2021-05-04
HUAZHONG AGRI UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, the traditional MassArray or SNaPshot system can only identify one or more known genotypes. For samples with specific genomic regions but unknown genotypes such as artificial mutagenesis and gene editing products, the current MassArray or SNaPshot The system cannot accurately obtain its genotype DNA sequence information

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  • A dna sequencing method for a specific region of the genome
  • A dna sequencing method for a specific region of the genome
  • A dna sequencing method for a specific region of the genome

Examples

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Embodiment 1

[0028] Example 1: Detection of Genotypes of Maize Genome Edited Plants

[0029] The use of CRISPP / Cas9 gene editing technology to genetically modify the crop genome can precisely change the functions of genes that control important traits, thereby achieving the improvement and optimization of crop traits. The edited recipient species needs to analyze the genome sequence of the target region to confirm the success of the editing, and at the same time, it is necessary to predict the changes in gene function and target traits based on the edited genotype sequence information. Therefore, identifying the edited sequence information of the target region of the target plant as early as possible is the key to affecting the efficiency of overall trait improvement.

[0030] Usually, gene editing technology can lead to the deletion, substitution or insertion of one or more bases in the target region. Because of the randomness of the base modification method, the genotype sequence inform...

Embodiment 2

[0061] Example 2: Sequence determination of polymorphisms in different germplasm maize genomes

[0062] Genome variation is the reason for the variation of species traits and the molecular basis for organisms to adapt to environmental changes. Studying the genome variation of different individuals within a species is an important research content for analyzing the molecular basis of individual trait variation, sorting out the evolutionary path of species, and discovering valuable genotypes. Usually, when the variation of a specific position in the genome of a species is known, its polymorphism can be determined using traditional Sanger sequencing or conventional MassArray, SNaPshot and other methods. However, if the polymorphic variation among different individuals is unknown, the above method cannot determine the variant sequence effectively or efficiently.

[0063] Using the method provided by the present invention, multiple extension primers can be designed to cover the sp...

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Abstract

The invention belongs to the field of biotechnology, in particular to a DNA sequencing method for a specific region of the genome. The invention designs multiple extension primers according to the reference genome sequence, and then uses the genome sequence to be tested as a template to extend the extension primer by one base, and finally assembles the DNA sequence of the detection region according to the measured extension product sequence. The invention can give full play to the advantages of high-throughput genotype analysis means such as MassArray flight mass spectrometry or SNaPshot, and is suitable for accurately determining DNA sequence information after natural or artificial mutations in known genome sequences. In addition, the method provided by the present invention can also be applied to the sequence determination of random and diverse gene editing products. This method can not only qualitatively detect whether the target region has been edited, but also accurately determine the edited sequence, and because Its multiple analysis feature makes the genotype analysis of the edited target region more efficient and accurate. Compared with the traditional Sanger sequencing, the invention improves the throughput, accuracy and speed of the sequencing work, and reduces the cost.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a DNA sequencing method for a specific region of the genome. Background technique [0002] Genome variation is the reason for the variation of species traits and the molecular basis for organisms to adapt to environmental changes. In addition, artificially induced genome variation (including radiation or chemical mutagenesis, gene editing, etc.) can produce new trait changes in species, so that excellent traits can be screened and solidified genetically, which has also become an important aspect of animal and plant breeding. basic process. Whether it is natural mutation or artificial mutation, the determination of the DNA sequence of a specific region has become the molecular basis for the study of trait variation and an important link in clarifying the molecular mechanism of variation. The traditional sequencing method is to obtain the mutated target region by methods such as PCR ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2533/101C12Q2537/143C12Q2531/113
Inventor 刘海军严建兵许洁婷
Owner HUAZHONG AGRI UNIV