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Gelsemium elegans Benth discriminating primer and Gelsemium elegans Benth discriminating PCR method

A technology of heartbroken grass and identification methods, applied in biochemical equipment and methods, microbe determination/testing, DNA/RNA fragments, etc.

Inactive Publication Date: 2015-05-20
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

Before the present invention was published, there was no publication or report on the PCR method and primers mentioned in this patent application

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  • Gelsemium elegans Benth discriminating primer and Gelsemium elegans Benth discriminating PCR method
  • Gelsemium elegans Benth discriminating primer and Gelsemium elegans Benth discriminating PCR method
  • Gelsemium elegans Benth discriminating primer and Gelsemium elegans Benth discriminating PCR method

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Embodiment Construction

[0016] 1 material

[0017] The materials used are shown in Table 1, including:

[0018] Table 1 Sample source table

[0019]

[0020] 2 Identification primer design

[0021] Select the universal primer psbA-trnH (trnH: 5'-CGC GCA TGG TGG ATT CAC AAT CC-3'psbA: 5'-GTT ATG CAT GAA CGT AAT GCT C-3') to amplify the samples of S. The psbA-trnH sequence of Lonicerae japonica in Genbank (Genbank accession number: GU1353313.1; JN045253.1), and then perform homologous alignment, and design a pair of specific primers F1 and F2 according to the variable region, the sequence is as follows:

[0022] F1: 5'-TCTCCGCCCTCTGCTCTA-3', F2: 5'-GGAAGAAAGGAGTTTTTTATGAGG-3'.

[0023] 3DNA extraction

[0024] 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 2.0mL microcentrifuge tube, add 900μL sterilized CTAB extract (2%CTAB, 100mmol / L Tris-HCl pH=8.0, 20mmol / L EDTA, 1.4mol / L NaCl), 0....

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Abstract

The invention belongs to the technical field of molecular biology, relates to the traditional Chinese medicine and traditional Chinese medicinal material identification field, and especially relates to a Gelsemium elegans Benth discriminating PCR amplification method and a Gelsemium elegans Benth discriminating specific primer. When the method is adopted to discriminate Gelsemium elegans Benth and honeysuckle flower, sample can be discriminated only through simple DNA extraction, PCR specific amplification and electrophoresis detection. The method has the advantages of simple operation, strong specificity and good repeatability, and is mainly used to rapidly identify Gelsemium elegans Benth and honeysuckle flower.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, relates to the field of identification of traditional Chinese medicines and Chinese medicinal materials, in particular to a PCR amplification method and specific primers capable of specifically identifying Herba chinensis. Background technique [0002] Gelsemium elegans is the stem vine of Gelsemium elegans (Gardn.el Champ) Benth. Also known as Hu vine, kudzu, water mangcao, big tea leaves, rotten intestine grass, it is a highly poisonous herb. It has the effects of expelling wind and attacking poison, reducing swelling and relieving pain. It can cure eczema, carbuncle, rheumatic arthralgia, etc. [0003] In recent years, poisoning accidents due to the mistaken consumption of S. chinensis have occurred in various places, and many studies have been carried out on it. It has been confirmed that the main components of the roots, stems, and leaves of S. wait. The investigation found tha...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q1/686C12Q2531/113C12Q2565/125
Inventor 黄璐琦袁媛李旻辉崔占虎蒋超
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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