Specific molecular marking primer and method for identifying sarcandra glabra and three adulterants

A labeled primer and specific technology, which is applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve the problem of low nuclear genomic DNA copy number, easy destruction of medicinal materials, and inability to identify molecular markers and other issues to achieve high efficiency

Active Publication Date: 2015-11-25
FUJIAN UNIV OF TRADITIONAL CHINESE MEDICINE
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Problems solved by technology

However, these genomic DNA-based molecular markers are susceptible to genomic DNA degradation, because nuclear genomic DNA has fewer copies than chloroplast DNA and is more likely to be destroyed in processed medicinal materials, making identification by its molecular markers impossible

Method used

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  • Specific molecular marking primer and method for identifying sarcandra glabra and three adulterants
  • Specific molecular marking primer and method for identifying sarcandra glabra and three adulterants

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Experimental program
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Embodiment Construction

[0015] instrument

[0016] PCR instrument (Eppendorf, model 5332), electrophoresis system (Beijing Liuyi Instrument Factory, model DYY-12), low-temperature refrigerated centrifuge (Eppendorf, model 5810R), gel imaging analyzer (BIO-RADChemiDocXRS), micropipetting machine (Eppendorf).

[0017] Reagent

[0018] 2×CTAB extract, 1×TAE buffer, agarose (Promega), ethidium bromide (Fluka), TransStart? TopTaq DNA Polymerase (Beijing Quanshijin Company), chloroform, absolute ethanol, isopropanol All are domestic analytically pure.

[0019] Material

[0020] The experimental samples in this study were identified and collected by Wei Yicong, a lecturer at Fujian University of Traditional Chinese Medicine, and collected from different origins: thirteen kinds of grass corals, four kinds of golden millet, four kinds of broad-leaf golden millet and three kinds of jellyfish were collected from different places in China. collect. The powders of five samples of Grass Coral and three ...

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Abstract

The invention relates to a specific molecular marking primer and a method for identifying sarcandra glabra and three adulterants, belonging to the technical field of biology. In the method, a multiple-PCR system is established: total amount is 20mu L, 10ng of DNA is extracted from a plant to be used as an amplification template, 1*TransStart TopTaq DNA polymerase is added, 2mu L of 10*TransStart TopTaq Buffer is added, 2.5mM of dNTP is added, 0.2mu M of downstream primer and three upstream specific primers SgF, CspF and CsF which are respectively 0.1mu M in amount are added, and the rest is sterile water. The method is used for molecule identification based on genetic background, and is not influenced by external factors such as production places of medicinal materials; and compared with a chemical method, the method disclosed by the invention is more accurate, and is simpler and more convenient.

Description

technical field [0001] The invention relates to a molecular-specific marker primer and a method for distinguishing coral coral from three kinds of obfuscated products, and belongs to the field of biotechnology. Background technique [0002] Sarcandraglabra (Thunb.) Nakai, commonly known as sarcandra, is usually used to treat tinea versicolor, purpura, rheumatic joint pain, and trauma. Modern studies have shown that it has pharmacological effects such as treating cancer, anti-tumor, anti-inflammation, anti-virus and non-specific immune enhancement. However, the germplasm resources of wild plants have been reduced in recent years. Because the appearance of Chrysanthemum praecox and Chrysanthemum broadleaf is very similar to that of Grass coral, it is easy to be confused with Grass coral. However, they have different properties and medicinal value, and these easily mixed plants also have corresponding hepatotoxic effects, and have been confirmed to have significant hepatotoxic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 魏艺聪车苏容梁一池
Owner FUJIAN UNIV OF TRADITIONAL CHINESE MEDICINE
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