Long-noded pit viper capillary electrophoresis DNA fingerprint spectrum and identification method

Abstract

The invention discloses and relates to a traditional Chinese medicine long-noded pit viper DNA identification technology. The method specifically comprises the following steps: extracting long-noded pit viper DNA by virtue of a CTAB process, and constructing a capillary electrophoresis fingerprint spectrum and an identification method. The method disclosed by the invention has the characteristicsof high resolution, simple operation, speediness and accurate results and can be used for identifying a long-noded pit viper sample.

Description

technical field [0001] The invention relates to identification technology of traditional Chinese medicinal materials, in particular to a viper capillary electrophoresis DNA fingerprint and identification method. Background technique [0002] Agkistrodon acutus (Guenther) is a dried body of Agkistrodon acutus (Guenther), an animal of Viperidae. It is more caught in summer and autumn. Cut open the abdomen of the snake, remove the viscera, wash, spread the abdomen with bamboo slices, and form a disc shape. After drying, remove the bamboo slices. It has the effects of expelling wind, dredging collaterals, and relieving spasm. For rheumatism obstinate numbness, numbness and spasm, hemiplegia, convulsions and convulsions, tetanus, leprosy, scabies. Driven by its economic interests, the drug varieties on the market are chaotic. [0003] At present, the identification methods of Viper mainly include character identification, microscopic identification and physical and chemical id...

AI Technical Summary

Problems solved by technology

[0006] 1. Poor repeatability and stability: Many factors will affect the amplification results of RAPD analysis, resulting in poor repeatability and stability of the analysis results

[0007] 2. Delayed detection technology: At present, the analysis of RAPD amplification products mainly adopts two methods: agarose gel electrophoresis and polyacrylamide gel electrophoresis

Polyacrylamide gel electrophoresis has high resolution and sensitivity, but the operation is cumbersome (including assembly, filling, electrophoresis, gel unloading, staining, etc.), time-consuming (the whole process takes hours to days), and the analysis efficiency is low. lower

Therefore, the current commonly used detection technology cannot meet the requirements of high resolution, high sensitivity, simplicity, and rapidity for the analysis of RAPD amplification products, and the lack of suitable detection methods has seriously restricted the application of RAPD technology in the construction of species DNA fingerprints.

[0008] 3. The identification method is unreasonable: when using RAPD technology to identify species at present, it is usually to select some representative amplified bands in the electrophoresis pattern, and identify them according to the presence or absence of these bands

Although this method makes the result analysis easier, it reduces the polymorphism of the amplification product and increases the chance of false positive results

Moreover, artificially selecting some amplified bands and discarding other bands also reduces the objectivity of the analysis results.

In addition, this identification method can only reflect the presence or absence of amplified bands, and cannot reflect the difference in the strength of the amplified bands.

[0009] Due to the above reasons, the current reports on the construction of species DNA fingerprints and species identification using RAPD technology have only theoretical research significance and cannot be applied in practice.

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