SNP (single nucleotide polymorphism) marker combination and identification method of small Meishan pig and raw meat product

An identification method and technology for meat products, which are applied in the field of SNP marker combination and identification of Xiaomeishan pigs and raw meat products, achieving the effect of simple and quick operation

Active Publication Date: 2018-02-16
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

[0003] In view of the above-mentioned deficiencies in the prior art, the present invention proposes a combination of SNP markers and an identification method for identifying Xiaomeishan pigs and raw meat products, using third-generation molecular marker identification and Sanger sequencing technology to identify X

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  • SNP (single nucleotide polymorphism) marker combination and identification method of small Meishan pig and raw meat product
  • SNP (single nucleotide polymorphism) marker combination and identification method of small Meishan pig and raw meat product
  • SNP (single nucleotide polymorphism) marker combination and identification method of small Meishan pig and raw meat product

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Embodiment Construction

[0054] Twenty ear tissue samples of Xiaomeishan pigs were selected for identification, and 10 ear tissue samples of each of the other twelve breeds (strains) were selected, and tissue DNA was extracted by the improved SDS method.

[0055] Using primer pairs C12, C6, C16, C71, and C79, PCR reaction amplification was performed.

[0056] The reaction system of PCR reaction is 10uL system: template DNA 1ng / uL, primer 1uL, H 2 O 3.8uL, 2×TaqPCR Masrer Mix 50uL; and / or, the reaction program of PCR reaction is pre-denaturation at 95°C for 2min, pre-denaturation at 95°C for 30s, annealing temperature at 60°C for 30s, extension at 72°C for 1min, cycle number 30 times, 72 °C for another 10 min.

[0057] Use 2% agarose gel, 1×TAE buffer as medium electrophoresis to detect the amplification result, and the gel electrophoresis conditions are current 10A, voltage 100 volts, time 40 minutes. The gel electrophoresis pattern of the amplification product of the C12 primer appears figure 1 Wh...

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Abstract

The invention provides an SNP (single nucleotide polymorphism) marker combination and identification method of a small Meishan pig and a raw meat product. The method comprises the following steps: extracting a genome DNA of raw pork or a meat product, performing AGE (agarose gel electrophoresis) and Sanger sequencing after performing PCR (polymerase chain reaction) amplification on the genome DNA,and identifying the small Meishan pig and the meat product thereof according to an SNP genotype of a characteristic site of a sequencing result, wherein in the step of Sanger sequencing, peculiar mutation occurs at identified sites: SNP6, SNP12, SNP16, SNP71 and SNP79. According to the method, the problem that an identification method of the small Meishan pig and the meat product thereof does notexist in the prior art is solved.

Description

technical field [0001] The invention relates to a technology in the field of food safety monitoring, in particular to a SNP marker combination and identification method for Xiaomeishan pigs and raw meat products. Background technique [0002] Meishan pig is one of the excellent local breeds (strains) in the Taihu Lake Basin in my country. It is famous for its high fecundity and delicious meat. With the development of sequencing technology, molecular markers have also developed from restriction fragment length polymorphism (RFLP) in the first generation, variable number tandem repeat polymorphism (SSR) in the second generation, to single nucleotide polymorphism ( SNP). Compared with the previous two generations of molecular markers, the third-generation molecular marker SNP has the advantages of rich variation, low requirement for DNA samples, high stability, accurate determination, simple detection method and high throughput. At present, the third-generation molecular mark...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q1/6888C12Q2600/156C12Q2535/101
Inventor 潘玉春岳阳王起山张哲徐忠孙浩刘登英赵伟马裴裴
Owner SHANGHAI JIAO TONG UNIV
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