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PCR method for rapid identification of fritillaria cirrhosa authenticity

A fritillary, authenticity technology, applied in the field of identification of Chinese medicinal materials, can solve problems such as hindering the large-scale promotion of fritillary molecular identification, long experimental time, heavy workload, etc., to shorten the detection time, improve molecular identification, accurate high degree of effect

Active Publication Date: 2018-07-10
GUIZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this way, when the PCR-RFLP method is used to identify Fritillaria sichuanensis, due to the existence of enzyme digestion steps, the workload is large, the experiment time is long and the cost is high, which hinders the large-scale promotion of the molecular identification of Fritillaria sichuanensis

Method used

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  • PCR method for rapid identification of fritillaria cirrhosa authenticity
  • PCR method for rapid identification of fritillaria cirrhosa authenticity
  • PCR method for rapid identification of fritillaria cirrhosa authenticity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Using the PCR-RFLP technology included in the Chinese Pharmacopoeia to identify the bases of Fritillaria sichuan, Fritillaria gansu, Fritillaria oleifera, Fritillaria wabu, Fritillaria saxa, and Fritillaria taibai

[0051] Step 1) Extraction of Genomic DNA

[0052] Six batches of each Fritillaria were taken, and the DNA was extracted using a DNA extraction kit.

[0053] ① Wash the samples with sterilized water, 75% ethanol and double distilled water in sequence, and place them on a clean bench to dry. After drying, grind it to a very fine powder with a mortar in an ultra-clean workbench, accurately weigh 0.1g of the sample powder, and transfer it to another mortar; Put the liquid into a 1.5mL centrifuge tube, and make up to 350μL with PBS buffer; ③Add 150μL Buffer C-L and mix well, then add 20μL Proteinase K, mix well, place in a 56℃ water bath for 30min, and mix up and down every 5min; ④Add 350μL Buffer P-D, mix well and centrifuge at 12000rpm for 10min; ⑤T...

Embodiment 2

[0067] Example 2 Utilizing the method described in the present invention, primer 1 is used to identify the original Fritillaria sichuanensis - Fritillaria sichuanensis, Fritillaria gansuensis, Fritillaria brunica, Fritillaria wabu, Fritillaria saxabilis and Fritillaria taibai

[0068] Step 1) Extraction of Genomic DNA

[0069] Six batches of each Fritillaria were taken, and the DNA was extracted using a DNA extraction kit.

[0070] According to the method of step 1) of Example 1, the genomic DNAs of Fritillaria sichuan, Fritillaria gansu, Fritillaria dark purple, Fritillaria wabu, Fritillaria sativa and Fritillaria taibai samples were extracted.

[0071] Step 2) PCR amplification

[0072] According to the method in step 2) of Example 1, the genomic DNA extracted in step 1) was used as a template, and PCR amplification was performed using primer 1.

[0073] Primer 1:

[0074] Upstream primer: 5'ACTATGCCCGCCCTACC 3',

[0075] Downstream primer: 5'GCTACGTTTCTTCATCGAT 3'.

[...

Embodiment 3

[0079] Example 3 Utilizing the method described in the present invention, primer 2 is used to identify the original Fritillaria sichuanensis - Fritillaria sichuanensis, Fritillaria gansuensis, Fritillaria nigra, Fritillaria wabu, Fritillaria saxabilis and Fritillaria taibai

[0080] Step 1) Extraction of Genomic DNA

[0081] Six batches of each Fritillaria were taken, and the DNA was extracted using a DNA extraction kit.

[0082] According to the method of step 1) of Example 1, the genomic DNAs of Fritillaria sichuan, Fritillaria gansu, Fritillaria dark purple, Fritillaria wabu, Fritillaria sativa and Fritillaria taibai samples were extracted.

[0083] Step 2) PCR amplification

[0084] According to the method in step 2) of Example 1, the genomic DNA extracted in step 1) was used as a template, and PCR amplification was performed using primer 2.

[0085] Primer 2:

[0086] Upstream primer: 5'ACTATGCCCGCCCTACC 3',

[0087] Downstream primer: 5'CTTCATCGATGCGAGAGC 3'.

[008...

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PUM

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Abstract

Belonging to the technical field of Chinese medicinal material identification, the invention relates to a PCR method for rapid identification of fritillaria cirrhosa authenticity. Specifically, the invention relates to primers for identification of fritillaria cirrhosa authenticity and application thereof in identification of fritillaria cirrhosa authenticity. The invention further relates to a PCR method for rapid and accurate identification of fritillaria cirrhosa authenticity with the primers.

Description

technical field [0001] The invention belongs to the technical field of identification of Chinese herbal medicines, and in particular relates to primers for identifying the authenticity of Fritillaria sichuanensis and its application in identifying the authenticity of Fritillaria sichuanensis; PCR method. Background technique [0002] The traditional Chinese medicine Chuan Fritillaria belongs to the Liliaceae Fritillaria plant, which is the dried bulb of Chuan Fritillaria, Gansu Fritillaria, Dark Purple Fritillaria, Wabu Fritillaria, Suosha Fritillaria or Taibai Fritillaria (National Pharmacopoeia Commission. People's Republic of China Pharmacopoeia: 2015 Edition. Part One [M]. China Medical Science and Technology Press, 2015), mainly produced in Sichuan and its neighboring provinces (Jiang Shunyuan, Sun Hongbing, Qin Jihong, etc., based on the function of growth suitability and quality suitability Research on regionalization of production [J], Chinese Journal of Traditional...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/683C12N15/11
CPCC12Q1/683C12Q1/686C12Q1/6895
Inventor 刘亭陆定艳杨友辉宋菲薛维娜李勇军何彬
Owner GUIZHOU MEDICAL UNIV
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