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Method for identifying beef

A beef and cytochrome technology, applied in the field of beef identification, can solve the problems of expensive and time-consuming reagents, consumables and testing equipment

Inactive Publication Date: 2017-01-04
宋胜辰
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In actual work, both traditional PCR and real-time quantitative PCR are used. The former usually takes a long time, while the latter has short detection time, but has the problem of expensive reagents, consumables and detection equipment.

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  • Method for identifying beef

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Embodiment Construction

[0019] Now in conjunction with embodiment, further illustrate the use of primer binding of the present invention Taq Good performance of DNA polymerase identification in beef. The present invention uses the Hangzhou Langji MG96+ type PCR instrument, but does not limit the use of instruments from other companies.

[0020] Prepare 4 identical groups, using the total DNA of beef / pork, pork liver and mutton as templates respectively, and other conditions are the same, a total of 4 tubes of 10 μL PCR reaction system: 10×Buffer: 1 μL; 10 mmol / L dNTPS: 0.4 μL; 0.5 μL each for upstream and downstream primers; template DNA: 0.5 μL; 5 U / μL Taq : 0.1 μL; 7 μL of sterilized pure water. The PCR program was as follows: 94°C denaturation / 25 s, 63°C annealing / 25 s, 72°C extension reaction / 25 s, 30 cycles. After the completion of PCR, 1.2% agarose gel electrophoresis was used to detect the experimental results.

[0021] The results showed that only one PCR product located in the middle po...

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Abstract

The invention relates to a method for identifying beef and relates to the technical field of food detection and animal molecular biologics. The method comprises the steps that a group of primers for specifically identifying a cytochrome gene b of cattle, the concentration of the total DNA of a sample purified with a phenol-chloroform extraction method is regulated to be 20 ng / microliters, the sample serves as a template for PCR detection, a 10-microliter PCR reaction system is prepared, and PCR is implemented according to a reaction procedure (modification is carried out at 94 DEG C for 25 s, annealing is carried out at 63 DEG C for 25 s, an extending reaction is carried out at 72 DEG C for 25 s, and operation circulates 30 times); 1.2% agarose gel is used for electrophoresis detection of a PCR product, and it is indicated that the sample is beef or contains bovine-derived ingredients if a 162 bp PCR product appears according to an electrophoresis detection result. Whether the sample is beef or not or whether beef ingredients are included in the sample or not can be quickly and accurately identified through the method, and the method can also be directly used for producing reagent kits for identifying beef.

Description

technical field [0001] The invention belongs to the technical field of food detection and animal molecular biology, relates to a method for identifying beef, and is suitable for identifying whether an unknown sample is beef or whether it contains beef components. Background technique [0002] With the improvement of the living standard of our country's residents, only the domestically produced meat is far from meeting the market demand, and a part of meat needs to be imported from abroad. These factors have created conditions for illegal traders at home and abroad to take advantage of the situation. In order to protect the legitimate rights and interests of consumers, it is necessary to establish an effective identification method for meat-derived ingredients. [0003] PCR (polymerase chain reaction), that is, DNA polymerase chain reaction, is a molecular biology technique invented in the 1980s that integrates high specificity, sensitivity, and high efficiency. It is widely...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q1/686C12Q2531/113C12Q2521/101
Inventor 宋胜辰杨仙玉李磊磊蒋桂瑾章胜楠李淑杰宋源普
Owner 宋胜辰