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PCR-RFLP (Polymerase chain reaction-restriction fragment length polymorphism) method for quickly identifying panax notoginseng and analogue panax stipuleanatus

A technology of Panax notoginseng and Wenshan, which is applied in the field of molecular identification of species, can solve the problems of sensory identification of the authenticity of Wenshan Sanqi raw materials, easy to mix into the screen side Sanqi, etc., and achieve the effect of overcoming the authenticity problem and accurate identification

Inactive Publication Date: 2013-06-26
YUNNAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0007] The present invention provides a reliable molecular identification method for Wenshan notoginseng and its easy-to-mix product, Pingbian notoginseng, aiming at the problems of difficult sensory identification of the authenticity of the raw material of Wenshan notoginseng and easy mixing of the raw material of Pingbian notoginseng

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  • PCR-RFLP (Polymerase chain reaction-restriction fragment length polymorphism) method for quickly identifying panax notoginseng and analogue panax stipuleanatus
  • PCR-RFLP (Polymerase chain reaction-restriction fragment length polymorphism) method for quickly identifying panax notoginseng and analogue panax stipuleanatus
  • PCR-RFLP (Polymerase chain reaction-restriction fragment length polymorphism) method for quickly identifying panax notoginseng and analogue panax stipuleanatus

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Embodiment Construction

[0015] Main instruments of the present invention: PCR instrument (CFX Manager), Hitachi desktop centrifuge, nucleic acid and protein analyzer (GeneQuant pro), water bath, refrigerator, electrophoresis instrument (DYY-6C), gel imaging analysis system (ImageQuant300). The following operations are conventional methods unless otherwise specified.

[0016] 1. Extraction of sample DNA

[0017] Wenshan notoginseng leaves and roots are collected from three-year-old plants in the Sanqi Science and Technology Demonstration Garden in Miao Township, Wenshan Zhuang and Miao Autonomous Prefecture, Yunnan Province; Pingbian notoginseng leaves and roots are collected from Wenshan notoginseng, Gulinqing Township, Maguan County, Wenshan Zhuang and Miao Autonomous Prefecture, Yunnan Province In the science and technology demonstration park, the leaves and roots of Wenshan notoginseng and Pingbian notoginseng were taken respectively, and DNA was extracted respectively. DNA was extracted accordin...

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Abstract

The invention discloses a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method for quickly identifying panax notoginseng and analogue panax stipuleanatus. According to the method, a primer adopted by PCR amplification in the method consists of an ITS (internal transcribed spacer) forward primer and an ITS reverse primer, the base sequence of the forward primer is as shown by SEQ ID NO:1, and the base sequence of the reverse primer is as shown by SEQ ID NO:1; and after a PCR amplification product is purified, a restriction enzyme MluC I and a restriction enzyme Nla IV are respectively used for carrying out enzyme digestion, and in an MluC I enzyme-digestion map and an Nla IV map, the enzyme-digestion maps of the panax notoginseng and the panax stipuleanatus are completely different, so that whether a sample is the panax notoginseng and whether the analogue panax stipuleanatus is mixed into the sample can be identified. By adopting the method, restriction enzyme enzyme-digestion maps of the panax notoginseng and the panax stipuleanatus are successfully established for the first time, the panax notoginseng and the panax stipuleanatus can be accurately and quickly identified, and the problem that sensory the authenticity of a panax notoginseng raw material cannot be easily identified through evaluation is solved.

Description

technical field [0001] The invention relates to the technical field of molecular identification of species, in particular to the molecular identification technology of restriction endonucleases (MluCI and NlaIV) on the authentic medicinal material Wenshan notoginseng of Yunnan Province and its easily mixed product Pingbian notoginseng. Background technique [0002] Wenshan Sanqi is the dried root of Panax notoginseng (Burk) F.H.Chen, a ginseng plant of Araliaceae, also known as Nanguo Shencao and Tianqi. It is one of the important and precious Chinese medicinal materials in my country. Panax notoginseng is called "gold is not exchanged". "Compendium of Materia Medica Supplements" records: "Ginseng is the first to nourish qi, and Sanqi is the first to nourish blood. It has the same taste and equal power, so it is called ginseng and notoginseng, and it is the most precious of Chinese medicines." Panax notoginseng and ginseng (Panax ginseng C.A.Mey.) both belong to the genus of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张广辉杨生超杨云陈军文李翠婷陈中坚朱书生何霞红
Owner YUNNAN AGRICULTURAL UNIVERSITY
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