Multiple PCR detection kit for kangaroos, guinea pigs and sewer rats and method

A detection kit and technology for Rattus norvegicus, which is applied to the multiplex PCR technology in the field of meat product detection, can solve the problems of expensive instruments and reagents, high requirements for detection technicians, long time consumption, etc., and achieve reliable detection results and simple identification methods. Effect

Pending Publication Date: 2020-05-08
BEIHUA UNIV
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  • Claims
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Problems solved by technology

However, the instruments and reagents used are expensive, time-consuming, require high tes

Method used

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  • Multiple PCR detection kit for kangaroos, guinea pigs and sewer rats and method
  • Multiple PCR detection kit for kangaroos, guinea pigs and sewer rats and method
  • Multiple PCR detection kit for kangaroos, guinea pigs and sewer rats and method

Examples

Experimental program
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Effect test

Embodiment 2

[0079] Example 2 Identification of baked kangaroo meat, guinea pig meat and Rattus norvegicus meat products

[0080] The materials were kangaroo meat purchased from imported supermarkets, postal kangaroo meat from Australia, farmed guinea pig meat, wild Rattus norvegicus (provided by Changchun Food and Drug Inspection Center), commercially available beef, and commercially available mutton (commercially available) after being baked in an oven at 100°C for 30 minutes. All samples were purchased from RT-Mart Supermarket in Jilin City).

[0081] 1. DNA extraction of the test product

[0082] (7) Take the meat sample, cut it into pieces about 0.05g, and put it in a centrifuge tube. Add 500 μL of P1 solution, 30 μL of P2 solution, and 15 μL of P3 solution, mix well, and shake in a 56°C water bath for 1 hour.

[0083] (8) Add 500 μL of P4 solution, mix for 5-10 minutes, and centrifuge at 10,000 r / min×5 minutes.

[0084] (9) Add the supernatant to an equal volume of P5 solution, and ...

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Abstract

The invention relates to the technical field of food detection and discloses a multiple PCR detection kit for kangaroos, guinea pigs and sewer rats. The multiple PCR detection kit is characterized bycomprising an animal tissue mitochondria DNA extraction system and an authenticating reaction system (being similar to a positive control solution reaction system and a negative control solution reaction system). An authenticating reaction total system is 50 microliters and contains 5 microliters of a reaction buffer solution, 2-5 microliters of dNTP, 23 microliters of MgCl, 1-5 microliters of TaqDNA polymerase, 2.25 microliters of a kangaroo primer, 3 microliters of a guinea pig primer, 2 microliters of a sewer rat primer, 2-6 microliters of DNA of a to-be-detected sample and the balance ofdouble distilled water. A multiple PCR authentication method for kangaroo meat, guinea pig meat and sewer rat meat comprises the steps of designing three specific oligonucleotide primers by virtue ofspecies specificity of cytochrome b, determining reaction procedures, determining a result, and the like; and the specificities of three rat meat can be simultaneously distinguished; and the method has the advantages of rapidness, reliable detection result and the like.

Description

technical field [0001] The invention relates to the application of multiplex PCR technology in the field of detection of meat products, more specifically, a DNA rapid identification kit and detection method for kangaroos, guinea pigs and Rattus norvegicus. Background technique [0002] Humans have long eaten rat meat. During the Tang Dynasty, the people in Lingnan liked to eat honey, "that is, the rats are naked and wriggling." Su Shi of the Northern Song Dynasty also ate rat meat in Lingnan and Hainan Island. There is a poem It goes: "You can see honey in the morning, and you can hear owls in your pillow at night." In some areas of China, there is a phenomenon of eating rat meat, mostly guinea pigs, bamboo rats, and kangaroos. The Rattus norvegicus is resolutely not allowed to appear on the human table. In 2013, there was a huge case of counterfeit mutton of brown rat meat in Jiangsu, China. In order to save costs, illegal traders purchased brown rat meat that had not been...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/686C12Q1/6888C12Q2600/16C12Q2600/166C12Q2537/143C12Q2545/113
Inventor 李明成段思琪
Owner BEIHUA UNIV
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