Molecular biology authentication method of bulbus fritillariae cirrhosae in Chinese patent medicine containing fritillariae
An identification method and a technology of Chinese patent medicine, which are applied in the field of molecular biology identification of Fritillaria sichuanensis, can solve the problems of high requirements and identification, and achieve the effects of less sampling, automation, and optimization of the PCR amplification reaction system
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Embodiment 1
[0028] In this example, twelve dried bulbs from the six base sources of Fritillaria cirrhosa and one dried bulb of the other four types of fritillary were used to prepare the powder of Shedan Fritillaria according to the composition ratio (6:1) of Fritillaria cirrhosa. As the test materials (see Table 1), genomic DNA was extracted, RAPD molecular marker analysis of designated primers was performed, and a standard fingerprint pattern was established with agarose gel electrophoresis bands for the identification of the components of the Chinese patent medicine Chuanbei.
[0029] The specific steps are as follows:
[0030] 1). After crushing each Snake Dan Fritillaria powder into very fine powder, it is then treated with TNE solution [200μmol / L tris-hydrochloric acid buffer, pH 8.0; 50μmol / L ethylenediaminetetraacetic acid, pH 8.0 and 250μmol / L Sodium Chloride] pretreatment, using improved CTAB method to extract genomic DNA. The operation is as follows:
[0031] (1) Weigh 0.1g of powd...
Embodiment 2
[0046] The test materials and operating steps of this example are the same as those in Example 1, wherein the random amplified polymorphic primer is S62GTGAGGCGTC, and a standard fingerprint is established for the identification of different base sources of the Chinese patent medicine Chuanbei.
[0047] figure 2 For the primer S62 (GTGAGGCGTC) to analyze the electrophoresis pattern of the RAPD labeling of sixteen Shedan Beimu San figure 1 the same. It can be seen that the primer can identify the source of each Fritillaria in Shedan Chuanbei San (the arrow shows the specific band of each Shedan Chuanbei San).
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