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DNA barcode based premier for identifying dinodon rufozonatum, PCR-RFLP method, and kit

A technology of PCR-RFLP and red chain snake, which is applied in the field of identification of traditional Chinese medicine varieties, can solve problems such as identification difficulties, achieve the effect of ensuring the quality of medicinal materials and shortening the operation time

Inactive Publication Date: 2014-11-26
SOUTHERN MEDICAL UNIVERSITY
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Problems solved by technology

At present, 50% of the money snake products on the market are counterfeit products from the red chain snake, and it is difficult to identify the two from the morphological characteristics

Method used

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  • DNA barcode based premier for identifying dinodon rufozonatum, PCR-RFLP method, and kit
  • DNA barcode based premier for identifying dinodon rufozonatum, PCR-RFLP method, and kit
  • DNA barcode based premier for identifying dinodon rufozonatum, PCR-RFLP method, and kit

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Embodiment Construction

[0024] The present invention will be described in further detail in conjunction with the following specific examples and accompanying drawings, and the protection content of the present invention is not limited to the following examples. Without departing from the spirit and scope of the inventive concept, changes and advantages conceivable by those skilled in the art are all included in the present invention, and the appended claims are the protection scope. The process, conditions, reagents, experimental methods, etc. for implementing the present invention are general knowledge and common knowledge in the art except for the content specifically mentioned below, and the present invention has no special limitation content.

[0025] 1. Materials

[0026] Six snake protozoa were collected mainly from Guangdong Province, Jiangxi Province, and Guangxi Zhuang Autonomous Region (Table 1). All samples were identified by Zhang Liang, South China Institute of Endangered Animals.

[0...

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Abstract

The invention belongs to the technical field of traditional Chinese medicine species identification, and provides a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identifying dinodon rufozonatum. The sequence of the premier is DK1-CO1:5'-CAA CTA ACC ACA AAG ACA TCG G-3', DK1-CO2:5'-CTT CTG GCT GGC CGA AAA ATC A-3'; and the characteristic DNA base sequence of dinodon rufozonatum is 5'-GTGCAC-3', which can be recognized and cut by restriction endonuclease APaL I. The identification comprises the following steps: extracting the DNA of a sample; using premiers DK1-CO1 and DK1-CO2 to carry out PCR amplification; using restriction endonuclease APaL I to digest the PCR products; carrying out agarose gel electrophoresis to obtain the analysis result; and comparing the analysis result with the PCR-RFLP characteristic result of dinodon rufozonatum to judge whether the provided sample is dinodon rufozonatum or not. The invention also provides a kit for the identification method. The identification is accurate, rapid, and reliable, and is capable of effectively identifying dinodon rufozonatum from other snakes.

Description

technical field [0001] The invention belongs to the technical field of identification of varieties of traditional Chinese medicines, and in particular relates to a primer, a PCR-RFLP method and a kit for identifying red chain snakes. Background technique [0002] Red chain snake (Dinodon rufozonatum) is a reptile of the genus Dinodonidae. It has anti-inflammatory, sedative hypnotic, and anti-convulsant effects. It is used for rheumatoid arthritis, detoxification and sores, general pain, chronic fistula, ulcers, and Ringworm etc. The red chain snake is a common counterfeit of the money white snake. With the increase in the price of medicinal materials and the gradual reduction of resources in recent years, more and more red chain snakes are being used as money snakes. At present, 50% of the money snake products on the market are counterfeit products from the red chain snake, and it is difficult to identify the two from the morphological characteristics. Finding a fast and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/683
Inventor 晁志李晓蕾曾伟萍
Owner SOUTHERN MEDICAL UNIVERSITY
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