Polymerase chain reaction (PCR) kit capable of rapidly detecting cervus derived products and preparation method

A deer product and kit technology, applied in the field of biological detection kits, achieves the effects of high speed, high sensitivity and simple operation

Inactive Publication Date: 2012-08-01
邢秀梅
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention provides a rapid detection method for detecting the authenticity of deer-derived deer products, which solves whether the deer products contain sika deer, red deer, Tarim red deer, red deer , Reindeer ingredients

Method used

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  • Polymerase chain reaction (PCR) kit capable of rapidly detecting cervus derived products and preparation method
  • Polymerase chain reaction (PCR) kit capable of rapidly detecting cervus derived products and preparation method
  • Polymerase chain reaction (PCR) kit capable of rapidly detecting cervus derived products and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: Composition and preparation of kit

[0047] 1) Preparation of DNA extraction solution

[0048] (1) Low-salt buffer (pH 7.6): Dissolve 1.21g Tris base (trishydroxymethylaminomethane) and 3.38g Na in 800ml distilled water 2 EDTA (disodium ethylenediaminetetraacetic acid), 0.76g MgCl 2 , 0.47gNaCl. Add concentrated hydrochloric acid to adjust the pH to the desired value, add distilled water to make up to 1L.

[0049] (2) Cell lysate: Add 1g SDS (sodium dodecyl sulfate) and 1ml Triton X-100 (polyethylene glycol p-isooctylphenyl ether) to 200ml low-salt buffer.

[0050] Extraction method: Add 200ul anticoagulant blood and 400ul low-salt buffer solution into a 1.5ml centrifuge tube, mix well, centrifuge at 1000RPM for 10min, discard the supernatant; add 500ul low-salt buffer solution, mix well, and centrifuge at 1000RPM for 5min at room temperature. Repeat once; carefully aspirate the supernatant, add 300ul of cell lysate, mix well, put in a water bath at ...

Embodiment 2

[0059] Example 2: Establishment of optimal reaction conditions for PCR.

[0060]For a PCR reaction, the most important reaction conditions are the annealing temperature and the extension temperature, so the optimal conditions for the PCR reaction are set from these two aspects. The setting of the optimal reaction conditions should first follow the principles of inter-species specificity and intra-species generality, and then select the temperature with the best amplification effect as the optimal reaction temperature of the primers on this basis.

[0061] 1) Extraction of DNA

[0062] Genomic DNA of sika deer was extracted from blood according to the method in Example 1.

[0063] 2) Preparation of PCR reaction system:

[0064] Add 1 ul of the DNA extracted in step 1) to the PCR reaction solution in Example 1 to form a 15 ul reaction system.

[0065] 3) Setting of optimal annealing temperature for PCR reaction.

[0066] The gradient range of annealing temperature was 54-6...

Embodiment 3

[0077] Embodiment 3: the specificity test of kit

[0078] 1) Demonstration that the primers are cross-species specific.

[0079] (1) In order to prove that the primers of the present invention have interspecies specificity, sika deer, red deer, Tarim red deer, red deer, reindeer, sambar, eld deer, white-lipped deer, elk, fallow deer, pig , cattle, sheep, and chicken blood genome DNA, the extracted DNA was used to prepare the PCR reaction system, and sterile water was used as a negative control.

[0080] (2) Preparation of PCR reaction system: 1 ul of DNA extracted in step (1) was added to the PCR reaction solution in Example 1 to form a 15 ul system.

[0081] (3) According to the reaction program: 94°C for 5min; 94°C for 30s, 56°C for 30s, 70°C for 30s, 70°C for 5min, 30 cycles of amplification. Perform PCR amplification.

[0082] (4) The result is as follows figure 1 : N: sika deer; C: red deer; Y: Tarim red deer; E: red deer; R: reindeer; 1: sambar; 2: Eld's deer; 3: ...

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Abstract

The invention discloses a PCR kit capable of rapidly detecting cervus derived products, which is used for identifying deer products rapidly and accurately. A kit is prepared by utilizing the PCR technology to amplify specific molecule fragments of each mitochondrial deoxyribonucleic acid (DNA) of sika deer, red deer, cervus elaphus yarkandensis, cervus elaphus and reindeer, is used for detecting whether a detected sample contains constituents of the deer, and can detecting mixed samples. The PCR kit capable of rapidly detecting the cervus derived products and a preparation method have the advantages of simple structure, high sensitivity, and fast speed.

Description

technical field : [0001] The invention discloses a PCR kit for quickly detecting the authenticity of deer-derived deer products, which is used for quickly and accurately identifying deer products, and belongs to the technical field of biological detection kits. Background technique: [0002] Deer products are some tissues or organs of deer, mainly including: deer antler, deer heart, deer kidney, deer fetus, deer tendon, deer whip, deer tail, venison meat and deer blood, etc. Since most deer products are precious Chinese medicinal materials and are expensive, it is common to see fake and inferior products pretending to be valuable deer products in the market. [0003] At present, the identification of deer products mostly adopts traditional methods, mainly including trait identification, microscopic identification and physical and chemical identification. Although the traditional identification method is simple, fast, and low in cost, there are many factors affecting the t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 邢秀梅杨福合査代明荣敏苏伟林吴琼
Owner 邢秀梅
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