Primer pair for identifying American ginseng and application of primer pair

A primer pair and American ginseng technology, applied in the field of molecular biology, can solve the problems of unfavorable application and promotion of rapid identification, cumbersome fluorescent staining methods, and long testing time

Inactive Publication Date: 2015-04-22
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] To sum up, different researchers used RAPD method, AP-PCR method, PCR-SSCP method, multiplex PCR method and other DNA molecular marker techniques to identify other medicinal materials in Panax ginseng, but these methods took a long time to test. , and both require electrophoresis detection to complete, so compared with the fluorescent staining method, it is cumbersome, which is not conducive to the application and promotion of on-site rapid identification

Method used

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  • Primer pair for identifying American ginseng and application of primer pair
  • Primer pair for identifying American ginseng and application of primer pair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, the preparation and use method of the kit for identifying American ginseng

[0042] 1. Design and synthesis of primer pairs for identification of American ginseng

[0043] In this example, the general primer matK (3F: 5'-CGTACAGTACTTTTTGTGTTTACGAG-3'1R: 5'-ACCCAGTCCATCTGGAAATCTTGGTTC-3') was selected to amplify the samples of Panax ginseng, Panax notoginseng, Panax notoginseng, Pearl ginseng and Panax notoginseng , sequenced, and then homologously compared the obtained sequences, and designed a pair of specific primers F1 and F2 according to the variable region, as a specific primer pair for identifying American ginseng. The sequence is as follows:

[0044] Primer F1: 5'-GGAAAATGCGGGTTATGACAGA-3' (SEQ ID NO: 1);

[0045] Primer F2: 5'-TTATGAGATTTGACTATCCCTTTGCTT-3' (SEQ ID NO: 2).

[0046] 2. Assembly of kits for identification of American ginseng

[0047] After the primers F1 and F2 designed and synthesized in step 1 are packaged separately, they are ...

Embodiment 2

[0060] Embodiment 2, using the kit prepared in embodiment 1 to identify American ginseng

[0061] Samples to be tested: 6 samples of Chinese herbal medicines shown in Table 1, Panax ginseng, Panax notoginseng, Bamboo ginseng, Pearl ginseng, and Panax notoginseng.

[0062] 1. Extract genomic DNA from the sample to be tested

[0063] Select about 0.03 g of dry medicinal material without mildew, put it in a pulverizer, grind it, and pass it through a 40-mesh sieve. Transfer the powder to a 2.0 mL microcentrifuge tube, add 900 μL of sterilized CTAB extract (recipe: 2% (2g / 100ml) CTAB, 100mmol / L Tris-HCl pH=8.0, 20mmol / L EDTA, 1.4mol / L NaCl), 0.02g PVP 40000, 10μL β-mercaptoethanol, fully vortexed and mixed, 65 ℃ water bath for 1.5h-2h, during which, shake gently 2-3 times. After completion, take it out and cool to room temperature, add 900 μL of chloroform-isoamyl alcohol (volume ratio 24:1), shake and mix well, and centrifuge at 12000 g for 10 min. Take the supernatant, add a...

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Abstract

The invention discloses a primer pair for identifying American ginseng and application of the primer pair. The primer pair for identifying or assisting the identification of the American ginseng, disclosed by the invention, is specifically a primer pair consisting of two single-chain DNA molecules as shown in sequence 1 and sequence 2 in a sequence table. Experiments prove that by adopting the primer provided by the invention, rapid and accurate distinguishment between the American ginseng and ginseng, pseudo-ginseng, panax japonicas, rhizoma panacis majoris and leatherleaf pseudo-ginseng can be realized by adopting a rapid PCR method and a fluorescence staining method; a fluorescent dye method is introduced for detecting real and false identification results, and a technical support is provided for realizing on-site application of identification of medicinal material molecules.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a pair of primers for identifying American ginseng and its application. Background technique [0002] The identification methods of Panax genus herbal medicinal materials mainly include morphological identification, physical and chemical identification, cell identification, etc. These traditional identification methods are easily affected by environmental conditions or limited by the plant itself, and there are many human interference factors, so they cannot directly reflect the genetics of the germplasm. characteristics, sometimes it is difficult to get accurate results (Zhou Yanqing. Application of DNA molecular marker technology in plant research. Beijing: Chemical Industry Press, 2005.). [0003] DNA molecular marker technology is a new detection technology that has emerged in the past two decades and has been continuously developed and improved. In recent years, DNA molecular...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895
Inventor 黄璐琦袁媛崔占虎蒋超
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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