American ginseng DNA detection reagent box and identification method

Abstract

The invention relates to the technical field of traditional Chinese medicine, in particular to an American ginseng DNA detection reagent box and an identification method. The American ginseng DNA detection reagent box comprises five parts, namely a sample pretreatment solution, a genome DNA extracting system, a PCR reaction system, a restriction enzyme reaction system and an observation result system. The identification method comprises six steps, namely detection sample pretreatment, genome DNA extraction, PCR primer design and synthesis, PCR reaction establishment, restriction enzyme reaction and result judgment. According to judgment standards, a 100 bp band and a 50 bp band occur simultaneously, the American ginseng is a salable product, and if only one band occurs or no band occurs, the American ginseng is a fake product. The PCR-AFLP identification method has the advantages of being simple, convenient, fast, reliable in detection result and the like, and the specificity of the American ginseng and the fake product can be identified accurately at the same time.

Description

technical field [0001] The invention relates to identification technology of Chinese medicinal materials, in particular to an American ginseng DNA detection kit and identification method. Background technique [0002] American ginseng (Panaxquinquefolius L.) is a perennial herb of the Araliaceae genus Panaxginseng C.A.Mey. American ginseng was originally a wild species, and later domesticated, and introduced and cultivated in countries all over the world. After 1975, my country successively introduced several batches of seeds from the United States, and the introduction was successful, among which the three northeastern provinces were the main planting areas. American ginseng is a precious nourishing medicinal material. Although it belongs to the same family as ginseng, there are great differences in efficacy and function. Although both have the effect of invigorating qi and promoting body fluid, American ginseng is cool in nature and sweet in taste, and its invigorating qi...

AI Technical Summary

Problems solved by technology

American ginseng is a precious nourishing medicinal material. Although it belongs to the same family as ginseng, there are great differences in efficacy and function. Although both have the effect of invigorating qi and promoting body fluid, American ginseng is cool in nature and sweet in taste, and its invigorating qi tends to nourish yin; Warm and bitter in nature, nourishing Qi tends to help Yang

The morphological characteristics of the two have many similarities, and it is easy to be confused if they are not carefully distinguished. In particular, it is difficult to accurately identify the Western ginseng cultivated in China based on the morphological characteristics. In actual work, we also encounter Western ginseng being confused and indistinguishable case

Song Qinxin et al. used ALM-ASA to identify single nucleotide polymorphism sites on the ITS genes of ginseng and American ginseng. This method designed specific primers for SNP sites and used adapter ligation-mediated allele-specific amplification. It has strong specificity and good stability, but it needs to design DNA adapters, the process is cumbersome, and it is difficult to popularize and apply it as a routine method [Song Qinxin, Zhang Xinyue, Bu Ying, et al.: ALM-ASA method to identify ginseng and American ginseng Single nucleotide polymorphism sites on ITS and 5.8s genes. Journal of China Pharmaceutical University, 2008, 39 (3): 274-278.]

Chen Ziyi and others used microsatellite markers to identify ginseng and American ginseng. This method uses SSR (simple repeat sequence) molecular markers as microsatellite DNA. It is necessary to design multiple pairs of primers for different SSR sites for amplification, and detect them by polyacrylamide gel electrophoresis. , although the accuracy is improved, but due to the large number of primers, the exploration of PCR reaction conditions and the detection of polyacrylamide gel electrophoresis are relatively time-consuming, and the mastery of experimental conditions also needs to be strictly controlled, so it is inconvenient in practical application. Chen Ziyi, Lu Xunan, Cheng Zhou, et al.: Application of microsatellite markers in the identification of ginseng and American ginseng. Journal of Fudan University (Natural Science Edition), 2011, 50(2): 185-191.]

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