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Amplification primer for detecting mink derived ingredients

A technology for amplification primers and water detection, which is applied in the field of amplification primers for detection of mink-derived components, and achieves the effect of species specificity and high sensitivity

Inactive Publication Date: 2014-01-08
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The RFLP method has its advantages, but it is necessary to select a low-cost, effective and suitable enzyme

Method used

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  • Amplification primer for detecting mink derived ingredients
  • Amplification primer for detecting mink derived ingredients
  • Amplification primer for detecting mink derived ingredients

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Establishment of the PCR detection kit and detection method of mink-derived component DNA

[0028] 1. Design of PCR detection primers for mink-derived DNA

[0029] From GenBank, 39 related mitochondrial sequences of carnivorous mustelids were retrieved, including mustelids, mink, otter, etc. This study refers to sable, mink bear, Japanese mink, blue fox, silver fox, raccoon dog, etc. published in GenBank For the mitochondrial genome sequence of animals, the sequences were compared by blast sequence. According to the conserved sequence of the species of sable and the analysis of specific sites, the primer design software primer5. Specific fragments of mink DNA are added, and no other animal DNA restriction fragments can be amplified.

[0030] 1. In the early stage of the experiment, based on the above principles, six pairs of primers were designed, named mink01 to mink06 respectively, and the designed primers were compared with the mitochondrial sequences of s...

Embodiment 2

[0064] Example 2 PCR detection method of mink DNA Specific detection experiment of mink-derived component DNA in meat and blood samples

[0065] In view of the phenomenon that mink meat is used as mutton rolls in the market, this test detects mink-derived components from the perspective of specific primer amplification fragments, in order to provide a detection method for detecting whether mink meat is adulterated in mutton rolls. It provides a molecular detection method for the quality inspection department. Mutton, beef, and chicken were purchased in the market and the genome was extracted for later use. DNA extraction from muscle tissue was done using the aforementioned kit. The detection kit and detection method containing specific primers of Example 1 were used to detect mink DNA samples. At the same time, the DNA samples of other fur animals such as blue fox and raccoon dog were detected to evaluate the specificity of this method.

[0066] The test results show that: ...

Embodiment 3

[0067] Example 3 The specificity experiment of the PCR detection method of mink-derived components to DNA in fur

[0068] Fur products are high-end consumer goods. Driven by profit, many people shoddy and even used fake artificial fur as mink fur, or used rabbit fur, dog skin, etc. as mink fur. Through further experimental means can be effectively identified and distinguished. The design of this experiment is as follows. Several kinds of animal fur were selected, and the DNA was extracted by the method of extracting genomic DNA from the fur. The genomic DNA of mink fur, fox fur, raccoon fur, dog skin and rabbit skin was respectively obtained, and the kit in Example 1 was used. And method for specific band test, the experimental results show that the above results can prove that the primers are only specific to mink.

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Abstract

The invention relates to an amplification primer for detecting mink derived ingredients and belongs to the detection of special economic animal mink derived ingredients. The amplification primer has nucleotide sequences that forward primers are 5'-CTTCAACCTCAACATCATCACC-3', and reverse primers are 5'-GACATACATTGTATTCATTCTAAGCG-3'. The simplicity, the species specificity and the high-degree sensitivity of a species specificity polymerase chain reaction (PCR) method are realized, and when the method is used, the detection of animal derived ingredients in food, feed and Chinese medical herbs is greatly improved and is very convenient. Compared with other molecular biology techniques, the amplification primer has the advantages that the price is lower, the speed is higher, and the amplification primer is more suitable for the ordinary detection of most samples.

Description

technical field [0001] The invention mainly relates to the detection of mink-derived components of special economic animals, in particular to specific primers and a kit for the mink gene detection method. Background technique [0002] Mink belongs to Mammalia, Carnivora, Mustelidae, and Mustela in animal taxonomy. In the wild, there are two species of American mink (Mestela vision) and European mink (Mustela lutreola). The minks that get fur are all descendants of American mink (Wang Jiyuan, 1990). my country's research on fur animal-related breeding technology began in the 1950s. Special economic animals have become an emerging characteristic industry in my country's agriculture because of their unique resource characteristics, important economic value and growing market demand. my country is a big country for raising special economic animals, and the number of fur animals alone reaches 328 million. Mink fur, fox fur and Persian lamb fur have long been the three pillar pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 李光玉孙伟丽赵家平张海华刘晗璐蒋清奎王夕国
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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