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Special primer, method and kit for identifying Meishan pig or meat product thereof

A meat product and kit technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve the effect of being beneficial to practical application, accurate results, and solving the problem of counterfeiting

Inactive Publication Date: 2015-05-06
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no method of identifying Meishan pig and its meat products by molecular identification methods at home and abroad. A method that can distinguish Meishan pig products from other varieties of pork products has been established to resolve counterfeit sales disputes involving Meishan pigs and their meat products. In order to effectively protect the market position and brand image of local Meishan pig breeds

Method used

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  • Special primer, method and kit for identifying Meishan pig or meat product thereof
  • Special primer, method and kit for identifying Meishan pig or meat product thereof
  • Special primer, method and kit for identifying Meishan pig or meat product thereof

Examples

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Embodiment 1

[0035] Example 1 Select the optimal annealing temperature when Meishan 1 is amplified

[0036] 1. Take 100mg of fresh meat from Meishan sow, put it into a grinder, add 1000μL of distilled water, grind for 3min, take 100μL of grinding solution, add 1mL of DNA extraction buffer, the extraction buffer is 10mmol / L Tris-HCl, 0.1mol / L EDTA, 0.5% SDS, and then add 25 μL of proteinase K to make the final concentration 100 μg / mL, mix well, and bathe in 50°C water for 3 hours. Extract once with an equal volume of phenol, collect the aqueous phase by centrifugation at 2500r / min, extract once with a mixture of equal volumes of phenol, chloroform and isoamyl alcohol, the volume ratio of the phenol, chloroform and isoamyl alcohol is 25:24 : 1, 2500r / min centrifugation collects aqueous phase; Extract once with the mixture of chloroform and isoamyl alcohol of equal volume, the volume ratio of described chloroform and isoamyl alcohol is 24:1; Add the LiCl of 5mol / L of equal volume , mix well,...

Embodiment 2

[0040] Example 2 Identification of Fresh Meat from Meishan Pig, Meidu Binary Miscellaneous and Du Dachang Sanyuan Miscellaneous

[0041] 1. Take 100mg of fresh meat from Meishan pig sow, Meidu binary mixed sow, Meishan pig boar and Meidu binary mixed boar, and frozen meat of Du Dachang Sanyuan mixed into the grinder and add 1000 μL Distilled water, grind for 5min, take 100μL grinding liquid, add 1mL DNA extraction buffer, described extraction buffer is 10mmol / L Tris-HCl, 0.1mol / L EDTA, 0.5% SDS, then add 25μL proteinase K again, make final concentration reach 100μg / mL, mix well, and bathe in 50℃ water for 3 hours. Extract once with an equal volume of phenol, collect the aqueous phase by centrifugation at 2500r / min, extract once with a mixture of equal volumes of phenol, chloroform and isoamyl alcohol, the volume ratio of the phenol, chloroform and isoamyl alcohol is 25:24 : 1, 2500r / min centrifugation collects aqueous phase; Extract once with the mixture of chloroform and iso...

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Abstract

The invention discloses a special primer, a method and a kit for identifying a Meishan pig or a meat product thereof. A sequence of the special primer is as shown in SEQ ID NO.1. The method comprises the following steps: (1), extracting genomic DNA (deoxyribonucleic acid) of fresh meat and / or the meat product; (2), taking genomic DNA in the step (1) as a template, and performing a PCR (polymerase chain reaction) by utilizing the primer with the nucleotide sequence as shown in SEQ ID NO.1. The method further comprises the following steps: (3), creating an electrophoretogram of amplified fragments in the PCR in the step (2); or (4), according to the electrophoretogram in the step (3), identifying the fresh meat and / or the meat product of the Meishan pig. The special primer and the method are utilized to effectively solve an identification problem of a Meishan pig variety and can distinguish the Meishan pig variety from other pig meat varieties or beef and mutton. The identification method is simple to carry out and accurate in result.

Description

technical field [0001] The invention relates to the field of biology, in particular to a special primer, method and kit for identifying Meishan pig or its meat products. Background technique [0002] Meishan pigs are originally produced in Jiading, Baoshan, Songjiang and other districts and counties of Shanghai in the Taihu Lake Basin, as well as Kunshan and Taicang in Jiangsu. characteristic. The Meishan pig breed in Jiading District has fresh and tender muscles, thin and dense muscle fibers, less water content, rich muscle fat, marble-shaped, commonly known as "pork belly", mellow flavor, delicate and soft. With the improvement of people's living standards, the quality requirements for livestock and poultry products are getting higher and higher. Many varieties such as Wai Sanyuan bred for the purpose of pursuing high yield are becoming less and less popular in China. Instead, pork market consumption pursues diversity. Refined, refined, and high-quality meat. Meishan po...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 史子学马志永张国平朱建忠王勃陈建生陆林根卫金良唐赛涌费琼慧于庆华魏建超刘珂李蓓蓓邵东华
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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