Detection kit for sheep origin component identification and detection of multi-species origin components in products

A detection kit, goat technology, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of high requirements, high cost, time-consuming and laborious, and achieve low cost and applicability. Broad and specific effect

Active Publication Date: 2016-02-03
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the fluorescence detection method has high requirements for equipment and is expensive; the enzyme digestion method is time-consuming and laborious; and the multiplex PCR is used to amplify the size of t...

Method used

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  • Detection kit for sheep origin component identification and detection of multi-species origin components in products
  • Detection kit for sheep origin component identification and detection of multi-species origin components in products
  • Detection kit for sheep origin component identification and detection of multi-species origin components in products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Specific detection of sheep-derived components in the sample of Example 1

[0077] 1. Sample preparation and storage

[0078] 1.1 Sampling

[0079] Collect 1 g of mutton samples and store them at -20°C for later use.

[0080] 1.2 DNA template preparation

[0081] The DNA template was prepared using the commonly used phenol-chloroform crude extraction method (Sambrook J, Fritsch EF, Maniartis T. Molecular Cloning Experiment Guide [M]. 2nd Edition. Jin Dongyan, Li Mengfeng. Beijing: Science Publishing Society, 1999.465-467) or recognized other extraction methods with the same efficacy, these methods are the commonly used methods reported.

[0082] 2. Primer Design

[0083] The DNA sequences of the primers in this embodiment are shown in Table 2 and SEQ ID NO: 2 and 3 in the sequence table.

[0084] Table 2 The sheep PCR amplification primers designed by the present invention

[0085]

[0086] The size of the expected amplified fragment is 175bp, and its nucleotide ...

Embodiment 2

[0108] Embodiment 2 sensitivity test

[0109] Using 0.1ng / μL, 1ng / μL, 10ng / μL, and 100ng / μL sheep DNA as templates, the nucleotide-specific fragment of SEQ ID NO.1 was amplified according to the conditions of Example 1. The result is as figure 2 As shown, there is amplification at a template concentration of 0.1ng / μL, indicating that the fragment amplified by the specific primer has good sensitivity.

Embodiment 3

[0110] Example 3 Detecting the primer set mixed with non-sheep-derived components in sheep-derived products

[0111] The present invention is for the detection of animal-derived components. Through the PCR amplification of the corresponding primers of each species, it is judged whether it is or contains the DNA of a certain species, and then it is judged whether the animal-derived components of a certain species are mixed.

[0112] 1 Extraction of DNA from samples

[0113] The extraction and storage methods of the DNA of each species are the same as those described in Example 1 above.

[0114] 2 Primer design and screening

[0115] In order to find the specific detection sequence of each species, we screen the specific sequence from the aspects of intra-species consistency and stability, inter-species specificity, copy number, etc., by comparing with the nucleic acid sequences of non-native species and different species of this species, The nucleotide sequences that are more...

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PUM

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Abstract

Belonging to the technical field of animal origin component molecular detection, the invention provides a detection kit for sheep origin component identification and detection of multi-species origin components in products. The kit includes primers shown as SEQ ID No.2&3, a reaction reagent, positive control and blank control. In addition, the kit also includes specific primers of ordinary cattle, buffalo, pigs, equus, rabbits, foxes, dogs, minks, rats, chickens and ducks, and the nucleotide sequences thereof are shown as SEQ ID No.4-25. The kit provided by the invention is employed for analysis to determine whether the sample contains sheep origin components and can effectively distinguish sheep and goats, and whether pigs, bos turus, buffalo, equus, rabbits, foxes, dogs, minks, rats, chickens, ducks and other animal ingredients are mixed. The invention provides an effective, accurate and reliable means for detection of the animal origin components in meat, hides and feed, and the kit and the method have the characteristics of simplicity, fastness, comprehensiveness, strong specificity, low cost, and wide applicability.

Description

technical field [0001] The invention relates to the field of animal molecular biology, in particular to the identification of sheep-derived ingredients and a detection kit for multi-species origin ingredients in products thereof. Background technique [0002] Mutton is one of the main meats eaten by the people of our country. It can not only keep out the wind and cold, but also nourish the body. It is a good tonic in winter. Its meat quality is similar to that of beef, but the meat flavor is stronger and delicious. Mutton is tender and rich in protein, with less fat and cholesterol than pork and beef. The content of minerals such as calcium, phosphorus, iron, potassium and iodine is higher than that of pork and beef, and has comprehensive nutrition. Li Shizhen said in "Compendium of Materia Medica": "Mutton can warm the middle and nourish the deficiency, nourish the middle and nourish the qi, appetize and keep fit, nourish the kidney qi, nourish the gallbladder and improve ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888
Inventor 刘榜付明王文君张静甄月然吴清清向胜男刘建建徐学文张庆德
Owner HUAZHONG AGRI UNIV
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