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Method for identifying beef

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

Inactive Publication Date: 2017-05-31
宋胜辰
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  • Summary
  • 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, 59°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 provides a method for identifying beef, and relates to the technical field of food detection and animal molecular biology. By designing a group of primers for specifically recognizing bovine mitochondrial genes, the total DNA (deoxyribonucleic acid) concentration of a sample purified by a phenol-chloroform extraction method is regulated to 20 ng / muL to be used as a PCR (polymerase chain reaction) detection template; a 10 mumL PCR reaction system is prepared; the PCR is performed according to the reaction procedure (degeneration for 25s at 94 DEG C, annealing for 25s at 59 DEG C, extension reaction for 25s at 75 DEG C, circulation for 30 times); a PCR product is detected through 1.2 percent agarose gel electrophoresis; according to the electrophoresis detection result, the generation of 178 bp PCR products shows that the sample is beef or the sample contains bovine derived ingredients. The method provided by the invention can be used for fast and accurately identifying whether the sample is the beef or the identification sample contains bovine derived ingredients or not. The method can also be directly used for producing a kit for identifying the 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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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q1/686C12Q2531/113C12Q2521/101C12Q2561/113C12Q2545/114
Inventor 宋胜辰杨仙玉章胜楠蒋桂瑾李磊磊李淑杰宋源普
Owner 宋胜辰