DNA barcode based primer for identifying bungarus multicinctus, PCR-RFLP method, and kit

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 bungarus multicinctus and counterfeits thereof. In the method, restriction endonuclease Spe I and BstE II are individually used to identify and cut the characteristic DNA base sequence of bungarus multicinctus so as to cut the COI segment of DNA code bar of bungarus multicinctus into three segments. The identification process comprises the following steps: extracting the DNA of a sample, using primers DK1-CO1 and DK1-CO2 to carry out PCR amplification; purifying the PCR products; using restriction endonuclease Spe I and BstE II to digest the PCR products; carrying out agarose gel electrophoresis to obtain the analysis result; and finally comparing the analysis result with the PCR-RFLP characteristic result of bungarus multicinctus to judge whether the provided sample is bungarus multicinctus or not. The invention also provides a kit for the identification method. The identification method is accurate, rapid, and reliable, and can be widely applied to identification of traditional Chinese herbals.

Description

technical field [0001] The invention relates to the technical field of variety identification of traditional Chinese medicines, in particular to a primer, a PCR-RFLP method and a kit for identifying white snakes and counterfeit products thereof. Background technique [0002] The golden snake is the dry body of the young snake of the Cobra family animal Bungarus multicinctus. Gas slightly fishy, ​​taste slightly salty. It has the effects of expelling wind, dredging collaterals and relieving spasm. For rheumatism obstinate numbness, numbness and spasm, apoplexy, squint, hemiplegia, convulsions, tetanus, leprosy, scabies, etc. The market of the golden white snake is called the small white snake. It is a rare and precious Chinese herbal medicine. However, with the deterioration of the ecological environment and the low production of the golden snake, the resources of its medicinal materials have decreased sharply. Due to the remarkable curative effect and shortage of medicine...

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Problems solved by technology

And it is difficult to make a single identification based on morphological characteristics, making the identification of varieties more and more difficult

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