Radix bupleuri capillary electrophoresis DNA (Deoxyribonucleic Acid) fingerprint spectrum and identification method

A technology of DNA fingerprinting and capillary electrophoresis, which is applied in the field of Bupleurum capillary electrophoresis DNA fingerprinting and identification, can solve the problems of reducing the objectivity of analysis results, poor repeatability and stability, and increasing false positive results, etc., to achieve easy automatic analysis and results Objective, Polymorphic Rich Effects

Inactive Publication Date: 2013-05-29
BEIHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] 1. Poor repeatability and stability;
[0011] 2. Although the amplification results are rich in polymorphisms, there is currently a lack of suitable detection methods;
[0012] 3. A large number of amplified bands make it difficult to analyze the results. At present, the commonly used method is to select repre

Method used

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  • Radix bupleuri capillary electrophoresis DNA (Deoxyribonucleic Acid) fingerprint spectrum and identification method
  • Radix bupleuri capillary electrophoresis DNA (Deoxyribonucleic Acid) fingerprint spectrum and identification method

Examples

Experimental program
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Example Embodiment

[0033] Example 1

[0034] Establishment of Capillary Electrophoresis DNA Fingerprint of Bupleurum

[0035] 1. DNA extraction of Bupleurum

[0036] Take 0.05~0.1g of Bupleurum (Bupleurum or Bupleurum angustifolia) control medicinal material, grind it thoroughly and transfer to a centrifuge tube, add 2×CTAB (1.4mol / L NaCl, 100mmol / L Tris pH 8.0, 20mmol / L EDTA) pH 8.0, 2% PVP, 2% CTAB, 3% β-mercaptoethanol) 200 μL, mix well, and keep in a water bath at 65°C for 1 hour with constant shaking during the period. Centrifuge at 12000r / min for 10min at 4℃. Aspirate the supernatant, add an equal volume of chloroform-isoamyl alcohol (24:1) for extraction 2 to 3 times, and centrifuge at 10000 r / min for 5 min. Take the supernatant and add 2 / 3 volume of absolute ethanol, mix well, and place at -20°C for 1 hour to precipitate nucleic acid. Centrifuge at 10000r / min for 5min at 4°C, discard the supernatant, and obtain DNA flocculent precipitate. The precipitate was rinsed twice with 80% ethanol a...

Example Embodiment

[0055] Example 2

[0056] Use the Bupleurum (Bupleurum) DNA fingerprint identification method of the present invention for the identification of a Bupleurum sample

[0057] Assuming that the existing batch of Bupleurum samples can not be determined whether they are real Bupleurum, therefore, it is identified according to the following method.

[0058] After the standard fingerprint of Bupleurum DNA is established, when a sample is to be identified, the mitochondrial DNA is first extracted by the CTAB method; the extracted DNA is used as a template, and the primers GTTACTCGAG and CCTGAATGCC are used for PCR amplification; Capillary electrophoresis analyzes and records the electrophoresis pattern to obtain the DNA fingerprint pattern of the batch of samples to be tested; use similarity analysis software to compare the similarity of the DNA fingerprint pattern of the sample to be tested with the DNA standard fingerprint pattern of Bupleurum spp., according to the similarity Identify th...

Example Embodiment

[0060] Example 3

[0061] Use the Bupleurum angustifolia DNA fingerprint identification method of the present invention for the identification of a certain Bupleurum angustifolia sample

[0062] Assuming that the existing batch of Bupleurum angustifolia samples cannot be determined whether they are true Bupleurum angustifolia, the DNA fingerprints of these samples are constructed according to the method of the present invention, and then similarity analysis software is used to compare the DNA fingerprints of the samples to be tested with The similarity of Bupleurum (Bupleurum angustifolia) DNA standard fingerprints is identified based on the similarity.

[0063] Identification result: If the similarity between the two is greater than 95% (including 95%), this batch of samples is true Bupleurum angustifolia; if the similarity between the two is less than 95%, then this batch of samples is not true Bupleurum angustifolia.

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Abstract

The invention relates to the technical field of traditional Chinese medicine identification and particularly relates to a radix bupleuri capillary electrophoresis DNA (Deoxyribonucleic Acid) fingerprint spectrum and an identification method. The radix bupleuri capillary electrophoresis DNA fingerprint spectrum comprises extraction of DNA, PCR (Polymerase Chain Reaction) amplification, analysis of an amplified product, and establishment of the radix bupleuri capillary electrophoresis DNA fingerprint spectrum. The identification method of a radix bupleuri DNA fingerprint comprises the steps of: establishment of the radix bupleuri capillary electrophoresis DNA fingerprint spectrum, establishment of a sample DNA fingerprint spectrum and result identification by utilizing similarity software. The fingerprint spectrum and the identification method can be used for identifying a radix bupleuri sample. The fingerprint spectrum and the identification method can be used for carrying out authenticity identification on radix bupleuri from heredity essence; and the method is rapid, accurate and objective and can be used for identifying the radix bupleuri sample.

Description

technical field [0001] The invention relates to identification technology of Chinese medicinal materials, in particular to a capillary electrophoresis DNA fingerprint and identification method of Bupleurum chinensis. Background technique [0002] Bupleurum is the root or whole plant of Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd., a perennial herb of Umbelliferae. The former is commonly called "Northern Bupleurum" and the latter is commonly called "South Bupleurum". Bupleurum bupleurum is a traditional precious Chinese herbal medicine in my country. It was first recorded in "Shen Nong's Materia Medica" and listed as top grade. "The Classic" says: Bupleurum "maintains confidantes and stomach stagnation, food accumulation, cold and heat pathogens, pushing old to bring about new." In modern times, it is used to disperse and reduce fever, soothe the liver and relieve stagnation, and promote sun and depression. Except for Hainan Province, Hong Kong and Macao Spe...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 苑广信李明成李洪宇王淼张丽华傅桂莲王冰梅
Owner BEIHUA UNIV
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