Loop-mediated isothermal amplification fluorescence detection method, primer and kit for identifying radix notoginseng ingredient, and application

A ring-mediated isothermal and amplification primer technology, which is applied in the field of warm amplification fluorescence detection, can solve the problems of the inconvenience of Sanqi and the high price of Sanqi, and achieve the effect of reducing the detection time, simplifying the detection process and reducing professional requirements

Inactive Publication Date: 2018-08-10
YUNNAN AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0003] Because Panax notoginseng is expensive, in order to make huge profits, many lawbreakers confuse real ones with inferior ones, causing inc

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  • Loop-mediated isothermal amplification fluorescence detection method, primer and kit for identifying radix notoginseng ingredient, and application
  • Loop-mediated isothermal amplification fluorescence detection method, primer and kit for identifying radix notoginseng ingredient, and application

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Embodiment Construction

[0066] The present invention will be further described in detail below in conjunction with the examples.

[0067]Those skilled in the art will understand that the following examples are only for illustrating the present invention and should not be considered as limiting the scope of the present invention. If no specific technique or condition is indicated in the examples, it shall be carried out according to the technique or condition described in the literature in this field or according to the product specification. The materials or equipment used are not indicated by the manufacturer, and they are all conventional products that can be obtained through purchase.

[0068] The PCR reaction concentrated solution that the present invention adopts is purchased from OptiGene Ltd., and the model is Isothermal MasterMix (IMM)

[0069] The plant genome extraction kit used in the present invention is TIANGEN plant genome extraction kit DP305.

[0070] Step (1), using the plant genom...

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Abstract

The invention relates to a loop-mediated isothermal amplification fluorescence detection method, primer and kit for identifying the radix notoginseng ingredient, and application, and belongs to the technical field of biological detection. The novel technology of fusing nucleic acid in vitro rapid amplification and fluorescence detection technologies is used; six specific primers are designed by aiming at target fragments; the whole isothermal amplification nucleic acid replication reaction process is monitored in real time through an amplification curve; the product is subjected to solubilitycurve analysis. A loop-mediated isothermal amplification method for identifying the radix notoginseng ingredient is built; the DNA level fast qualitative detection on whether the radix notoginseng iscontained in a sample to be tested or not is performed; the detection use time is greatly shortened; the fast detection of the radix notoginseng ingredient is realized; the computer experiment can beperformed without purification after DNA extraction; the six primers correspond to the recognition of eight specific regions of a target sequence, so that high-degree specificity of the detection reaction is ensured; good application prospects are realized.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a ring-mediated isothermal amplification fluorescence detection method, primers, kit and application for identifying components of Panax notoginseng. Background technique [0002] Panax notoginseng is a traditional precious Chinese medicinal material in my country, one of the 40 kinds of bulk medicinal materials. It has a history of more than 400 years of application and has a long-standing reputation at home and abroad. Sanqi has been known as the "hemostatic medicine" since ancient times. Due to the unique and obvious effects of Panax notoginseng, more and more people began to take it in their daily life; there are more than 360 kinds of Chinese patent medicines using it as raw materials, involving more than 1,350 pharmaceutical companies; its price remains high, and price fluctuations are enough to affect the Thousands of pharmaceutical companies. Wha...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/6895C12Q2531/119C12Q2563/107
Inventor 卢迎春贾兵张广辉陈军文杨生超宋婉玲朱书生朱有勇陈中坚魏富刚张强皓韦海南
Owner YUNNAN AGRICULTURAL UNIVERSITY
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