American ginseng DNA detection reagent box and identification method

A detection kit and identification method technology, applied in the field of identification of traditional Chinese medicinal materials, can solve problems such as inconvenience in practical application, difficulty in popularization and application, differences in drug efficacy and function, and achieve reliable detection results

Inactive Publication Date: 2015-11-11
BEIHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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

Method used

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  • American ginseng DNA detection reagent box and identification method
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  • American ginseng DNA detection reagent box and identification method

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Embodiment 1

[0069] 1. Pretreatment of test samples

[0070] Test samples (two types, 1 is authentic American ginseng, 2 is ginseng, American ginseng counterfeit, both provided and identified by the China Institute for the Control of Pharmaceutical and Biological Products), each take 0.03g, rinse with pre-treatment solution after scrubbing, and dry at room temperature , irradiate with ultraviolet light for more than 30 minutes, use scissors to cut the sample to about 1-2mm, put it in a centrifuge tube, and weigh it for later use;

[0071] 2. Genomic DNA Extraction

[0072] (a) Sample lysis Add 1ml of lysate to the pretreated sample, place it in a water bath, bathe at 65°C for 30 minutes, and rotate at 100r / min;

[0073] (b) After precipitation and lysis, centrifuge the sample at 13000r / min at 4°C for 10min, absorb the supernatant, add an equal volume of precipitation solution, mix well, centrifuge at 13000r / min at 4°C for 2min, discard the supernatant, and keep the precipitate;

[0074] (c...

Embodiment 2

[0093] Example 2 Identification of commercially available American ginseng medicinal materials

[0094] 1. Materials

[0095] 5 samples of commercially available American ginseng, 1 genuine product of American ginseng (provided and identified by Jilin Drug Control Institute)

[0096] 2. Method

[0097] 2.1 Pretreatment of test samples Take 0.03g of each test sample, scrub it clean, rinse with pretreatment solution, dry at room temperature, irradiate with ultraviolet light for more than 30min, use scissors to cut the sample to about 1-2mm, and place it in a centrifuge tube , weighed for use;

[0098] 2.2 Genomic DNA extraction

[0099] (a) Sample lysis Add 1ml of lysate to the pretreated sample, place it in a water bath, bathe at 65°C for 30 minutes, and rotate at 100r / min;

[0100] (b) After precipitation and lysis, centrifuge the sample at 13000r / min at 4°C for 10min, absorb the supernatant, add an equal volume of precipitation solution, mix well, centrifuge at 13000r / m...

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2521/301C12Q2535/138
Inventor 王雪松李明成艾金霞夏薇张丽华高丽君
Owner BEIHUA UNIV
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