Chinese-medicine Fritillaria-thunbergii ploymerase chain reaction appraising trigger and appraising method

A technology of fritillary and polymerase, which is applied in the identification of primers and identification by polymerase chain reaction of fritillary in traditional Chinese medicine, and can solve problems such as different

Inactive Publication Date: 2003-08-27
CHINA PHARM UNIV
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Problems solved by technology

However, this method does not have a unified method and primers for all Chinese medicinal materials. For different Chinese medicinal materials, the identification methods and primers are different.

Method used

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Embodiment Construction

[0030] Firstly, the total DNA was extracted from the original plants of different species of fritillaria, multiple gene fragments were amplified by PCR method and purified separately, DNA sequence analysis was performed on each purified fragment, and fritillary fritillary and its related species, mixed species were established. DNA sequence database, on the basis of comparing databases, select appropriate DNA fragments, and design a pair of differential PCR primers for the occurrence of counterfeit and mixed medicinal materials:

[0031] Primer 1: 5'-CTAAATAGGCGG GCGAGCTAATC-3'

[0032] Primer 2: 5'-TATTAGCATTAATCGATCGAATTC-3'

[0033] Use an automatic DNA synthesizer to synthesize according to the DNA sequence of the above-mentioned identification primer, and then the white powder of the identification primer can be obtained without crystallization.

[0034] The above primers were used to identify the authenticity of Fritillaria fritillata. Taking the reaction system 30 μl a...

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Abstract

The invention belongs to traditional Chinese medicine appraisal field, it is priming for appraising the Chinese Fritillaria taipaiensis. The priming DNA sequence is: the priming 1:5'-CTAAATAGGCGGGCGAGCTA ATC-3'; the priming 2:5'-TATTAGCATT AATCGATCGAATTC-3'. The appraising method is: extracts the total DNA of medicinal materials; uses a couple of priming AS, AS-1 to expand the DNA in order to check its quality, if there is positive expansion, the model DNA is eligible; uses the appraising priming to expand it; uses electrophoresis to check the result, if gets the positive expansion, it is Chinese Fritillaria taipaiensis, if not, it isn't zhebeimu.

Description

1. Technical field: [0001] The present invention is a kind of primer and its identification method for identification of traditional Chinese medicine Fritillaria japonicus, relating to the identification of Chinese medicinal materials [0002] technology field. 2. Background technology: [0003] Fritillaria is an important class of antitussive and expectorant drugs, and its active ingredients are Fritillaria alkaloids. The 2000 edition of the Chinese Pharmacopoeia recorded 9 species of Fritillaria. Different species of Fritillaria have different types and contents of alkaloids, as well as differences in their toxicity and efficacy. Fritillaria fritillaria is currently the most widely used in the market, but it is also highly toxic, and its dosage should be controlled when using it. Therefore, it is extremely important to accurately identify Fritillaria fritillata. However, the use of morphological and physical and chemical methods for its identification requires extensive ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/25C12Q1/68
Inventor 李萍王冲之彭昕
Owner CHINA PHARM UNIV
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