A high-throughput and low-cost SNP typing method based on the principle of liquid-phase molecular hybridization

A molecular hybridization and typing method technology, applied in the field of molecular biology DNA genetic markers, can solve the problems of inability to achieve multi-sample high-throughput typing, and achieve low typing cost, low cost, simple and controllable process steps. Effect

Active Publication Date: 2015-12-30
OCEAN UNIV OF CHINA
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Problems solved by technology

However, for the fixed-point typing of known SNP sites, the existing sequencing typing method does not give full play to the advantages of high-throughput sequencing, and it is still a low-to-medium throughput typing method, which cannot realize the simultaneous analysis of multiple sites and multiple samples. high-throughput typing

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  • A high-throughput and low-cost SNP typing method based on the principle of liquid-phase molecular hybridization
  • A high-throughput and low-cost SNP typing method based on the principle of liquid-phase molecular hybridization
  • A high-throughput and low-cost SNP typing method based on the principle of liquid-phase molecular hybridization

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Embodiment 1

[0045] The present invention provides MuLTISNP (Multi-Locus Targeted Indeland SNP genotyping), a novel SNP typing technology based on a high-throughput sequencing platform, and uses Genomic DNA of Chlamys farreri as an example for illustration. like figure 1 As shown, this embodiment takes 3 Chlamys farreri individuals, 1296 SNP sites, and each individual as an example to repeat the experiment twice, and describes the technical solution of the present invention in detail through the implementation of the experiment.

[0046] The high-throughput and low-cost SNP typing method based on the liquid-phase molecular hybridization principle of the present invention

[0047] MultiSNP specifically includes the following steps:

[0048] 1. Extract scallop genomic DNA and carry out biotin labeling, the method is as follows:

[0049] 1) Extract genomic DNA from Chlamys farreri: Take about 0.1 g of adductor muscle from Chlamys farreri, add 500 μl of STE lysis buffer (NaCl: 100 mM; EDTA: ...

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Abstract

The invention provides a high-throughput low-cost SNP (single nucleotide polymorphism) genotyping method based on a liquid molecular hybridization principle. The method comprises the following steps: extracting biological genome DNA (deoxyribonucleic acid) and carrying out biotin labeling on the biological genome DNA; designing site-specific hybridization primers LSP1 and LSP2 and carrying out 5' phosphorylation on LSP2; hybridizing an LSP1 mixture and an LSP2 mixture with the genome DNA to obtain a hybridization adsorption product; carrying out extended linkage reaction to obtain a target DNA fragment; carrying out a round of PCR (polymerase chain reaction) amplification on a universal primer; carrying out PCR amplification on the Barcode specific primer of the recovered target fragment; carrying out high-throughput sequencing; obtaining SNP site genotyping information through analysis. The method combines the site selection flexibility of the liquid hybridization technology with the advantages of high throughput and low cost of the high-throughput sequencing technology and has great application value and wide popularization prospect in the research fields such as large-scale screening of SNP, genome-wide association study, population diversity evaluation, gene function analysis and the like.

Description

technical field [0001] The invention belongs to the technical field of DNA genetic markers in molecular biology, and in particular relates to a high-throughput and low-cost SNP typing method based on the principle of liquid-phase molecular hybridization. Background technique [0002] Single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP) mainly refers to the DNA sequence polymorphism caused by single nucleotide variation at the genome level. SNP has the advantages of wide distribution, huge number, rich information, and easy detection. It has become the third generation of genetic markers after RFLP and SSR. At present, SNP is widely used in the construction of genetic maps, population genetics and kinship, association analysis and gene function analysis and other fields. [0003] At present, for the typing of known SNP sites, many detection methods have been established according to different research needs and experimental conditions. The main analysis princ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6827C12Q1/6869C12Q2563/185C12Q2535/122
Inventor 包振民王师焦文倩窦锦壮于茜荀小罡张玲玲胡晓丽陆维
Owner OCEAN UNIV OF CHINA
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