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Bupleurum DNA (deoxyribonucleic acid) identifying kit and identifying method

An identification method and kit technology, applied in the direction of biochemical equipment and methods, microbiological determination/inspection, etc., can solve the instability of the method, the lack of practicability of the research results, and the lack of Bupleurum molecular markers in the dried roots of Bupleurum medicinal materials, etc. question

Inactive Publication Date: 2011-09-21
BEIHUA UNIV
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Problems solved by technology

Most of the research is on the young tissues of Bupleurum chinensis (mostly fresh or dried leaves), and the medicinal part is the dried root of the medicinal material that contains a large number of secondary metabolites, and is usually processed and concocted, so the research results lack practicability; and Some studies on the dried roots of Bupleurum have not obtained the specific molecular markers of Bupleurum, and the method is not stable

Method used

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  • Bupleurum DNA (deoxyribonucleic acid) identifying kit and identifying method

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Embodiment Construction

[0035] The present invention will be further described below using examples.

[0036] A Bupleurum DNA identification kit, which comprises:

[0037] (1) DNA extraction reagent: the total system is 1000 μl, 2% CTAB, 100 mmol / L Tris-HCl, 30 mmol / L EDTA, 2.5 mmol / L NaCI, 3% PVP-40.

[0038] (2) Bupleurum identification system, including three parts:

[0039] ①Bupleurum identification system: the total system is 100μl, containing 10μl buffer, 12.5mM dNTP 4-10μl, 0.1mM "primer 1" 1.5-4.0μl, Taq DNA polymerase 2-10μl, sample DNA to be tested 2-5μl, and the rest for double distilled water.

[0040] ②Bupleurum positive control system: the total system is 100μl, containing buffer 10μl, 12.5mM dNTP4-10μl, 0.1mM "Primer 1" 1.5-4.0μl, Taq DNA polymerase 2-10μl, Bupleurum DNA 2-5μl, the rest is double distilled water.

[0041] ③Bupleurum negative control system: the total system is 100μl, containing buffer 10μl, 12.5mMdNTP 4-10μl, 0.1mM "Primer 1" 1.5-4.0μl, Taq DNA polymerase 2-10μl, b...

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Abstract

The invention relates to the technical field of traditional Chinese medicine detection, in particular to a bupleurum DNA (deoxyribonucleic acid) identifying kit and identifying method. The identifying kit comprises a DNA extraction reagent, a bupleurum identifying system and a bupleurum scorzonerifolium willd identifying system. The bupleurum DNA identifying method comprises the steps of: designing two pairs of specific oligonucleotide primers of the bupleurum ribosomal DNA, artificially synthesizing two pairs of primers, establishing a bupleurum DNA identifying kit, determining a reaction process, judging the result, and the like. The bupleurum DNA identifying kit and identifying method can be used for identifying and detecting traditional Chinese medicines, the bupleurum DNA identifying kit can accurately identify the specificity of bupleurum and can simultaneously identify the bupleurum, the bupleurum scorzonerifolium willd and confusing species thereof, and the identifying method has the advantages of simplicity, quickness and the like.

Description

technical field [0001] The invention relates to a DNA identification kit and an identification method for identification of Bupleurum (Bupleurum and Bupleurum angustifolia) and its easily mixed varieties using biotechnology. Background technique [0002] Bupleurum bupleurum is a perennial herb of the genus Bupleurum in the Umbelliferae family, and its roots and whole plants are used as medicine. The function is to harmonize the interior, soothe the liver and relieve stagnation, elevate the middle-qi, etc. It is a commonly used traditional Chinese medicine in clinical practice. There are 36 species of Bupleurum genus plants in my country, 17 varieties, and 7 varieties. The Pharmacopoeia of the People's Republic of China (2005 Edition) only stipulates that Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. are genuine medicines. However, there are more than a dozen mixed varieties on the market. [0003] Especially in recent years, due to the serious reduction of re...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张丽华李明成苑广信王冰梅
Owner BEIHUA UNIV
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