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Molecular-specific marker primers and methods for distinguishing Grass coral and 3 kinds of confounding products

A labeled primer and specific technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problems of small number of nuclear genome DNA copies, inability to identify with molecular marker technology, easy damage of medicinal materials, etc., and achieve high efficiency.

Active Publication Date: 2017-11-17
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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  • Molecular-specific marker primers and methods for distinguishing Grass coral and 3 kinds of confounding products
  • Molecular-specific marker primers and methods for distinguishing Grass coral and 3 kinds of confounding products

Examples

Experimental program
Comparison scheme
Effect test

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-RAD ChemiDoc XRS), trace Pipettes (Eppendorf).

[0017] Reagent

[0018] 2×CTAB extract, 1×TAE buffer, agarose (Promega Company), ethidium bromide (Fluka Company), TransStart®TopTaq DNA Polymerase (Beijing Quanshijin Company), chloroform, absolute ethanol, iso All propanol is of domestic analytical grade.

[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 t...

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Abstract

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. Construct a multiplex PCR system: the total amount is 20 μL, extract 10 ng of DNA from plants as an amplification template, add 1×TransStart TopTaq DNA polymerase, 2 μL 10×TransStart TopTaq Buffer, dNTP 2.5mM, 0.2 μM downstream primer trnL‑Ff And the three upstream specific primers SgF, CspF and CsF are 0.1 μM each, and the rest are sterile water. This method is based on the genetic background for molecular identification, and is not affected by external factors such as the place of origin of the medicinal material. Compared with the chemical method, it is more accurate, 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] Sarcandra glabra (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 hepatotoxi...

Claims

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Application Information

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Patent Type & Authority Patents(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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