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Hangzhou white chrysanthemum PCR identification kit and application thereof

A kit, the technology of Chrysanthemum chrysanthemum, applied in the field of DNA molecular identification, can solve the problems of difficulty in distinguishing traditional Chinese medicine Chrysanthemum chrysanthemum, and achieve the effects of short time consumption, strengthening the quality supervision of traditional Chinese medicine, and good repeatability

Active Publication Date: 2022-03-01
ZHEJIANG UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a PCR identification kit and detection method capable of accurately identifying the traditional Chinese medicine Chrysanthemum chrysanthemum, including PCR primers, templates and detection steps of the key reagents of the PCR detection method, which solves the problem that the traditional Chinese medicine Chrysanthemum chrysanthemum is difficult to distinguish

Method used

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  • Hangzhou white chrysanthemum PCR identification kit and application thereof
  • Hangzhou white chrysanthemum PCR identification kit and application thereof
  • Hangzhou white chrysanthemum PCR identification kit and application thereof

Examples

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Effect test

Embodiment 1

[0025] 1. Acquisition of PCR primers for the key reagents of Chrysanthemum chrysanthemum PCR detection method

[0026] Genomic DNA was extracted from Hangbaiju (Tongxiang, Zhejiang), Huaiju (Jiaozuo, Henan), Chuzhou (Chuzhou, Anhui) and Boju (Bozhou, Anhui) using Plant DNA Rapid Extraction Kit (Shanghai Sangon Biological Co., Ltd.). Using 5000 random primers, RAPD analysis was carried out on four kinds of medicinal chrysanthemums by RAPD amplification method, and a specific amplification band of Chrysanthemum chrysanthemum was screened. There is no medicinal chrysanthemum, and the DNA molecule in this band is the specific DNA molecular marker of Chrysanthemum chrysanthemum. The Chrysanthemum chrysanthemum-specific DNA molecular marker was recovered from the agarose gel, cloned, and sequenced. The sequencing result is shown in SEQ ID NO.1 and named HBJ893.

[0027] According to the sequence of HBJ893, design a pair of specific primers HBJ893-1 and HBJ893-2, the primer sequence...

Embodiment 2

[0042] 1. Study on the Accuracy of Chrysanthemum Chrysanthemum PCR Detection Method

[0043] From Hangbaiju (Tongxiang, Zhejiang), Huaiju (Jiaozuo, Henan), Chuju (Chuzhou, Anhui), Boju (Bozhou, Anhui), wild chrysanthemum, dandelion, Zhebaizhu, Anbaizhu, Zhebai, Anbai, Shanbai , Genomic DNA extracted from Radix Paeoniae Alba plants. Using the genomic DNA of the above 12 materials as a template, PCR amplification was performed using the specific primer "HBJ893-1 / HBJ893-2". The PCR reaction solution, amplification procedure, and detection method were the same as in Example 1, except that the DNA template was replaced to verify The accuracy and reliability of the Chrysanthemum chrysanthemum PCR detection method, see the results figure 1 As shown, lanes 1 to 12 are Hangbaiju (Tongxiang, Zhejiang), Huaiju (Jiaozuo, Henan), Chuju (Chuzhou, Anhui), Boju (Bozhou, Anhui), wild chrysanthemum, dandelion, Zhebaizhu, Anbaizhu, Zhebai , Anbai, Shanbai, Chuanbai, lane M is the marker.

[0...

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Abstract

The invention discloses a Hangzhou white chrysanthemum PCR (Polymerase Chain Reaction) identification kit and application thereof. Specific primers in the kit are HBJ893-1 and HBJ893-2. The invention provides a method for rapidly and accurately identifying the Hangzhou white chrysanthemum, can be widely applied to variety identification in various links of seed selection and breeding, planting, harvesting and storage, selling and clinical application of the traditional Chinese medicine of the Hangzhou white chrysanthemum, and has great significance in identifying the authenticity of the traditional Chinese medicine, fighting against counterfeit products, enhancing the quality supervision of the traditional Chinese medicine and promoting the modernization development of the traditional Chinese medicine.

Description

(1) Technical field [0001] The invention relates to a DNA molecular identification method, in particular to a PCR identification kit, a specific primer, a template and an identification method of the traditional Chinese medicine Chrysanthemum chrysanthemum. (2) Background technology [0002] Chrysanthemum is the dry head of Chrysanthemum chrysanthemum (Chrysanthemum morifolium Ramat.). The varieties of medicinal chrysanthemum include Hangbaiju, Huaiju, Chuju, Boju, and Gongju. Chrysanthemum contains active ingredients such as volatile oil, inulin, adenine, choline, amino acid, flavonoids, phenolic acids and locust, which have the functions of antibacterial, antiviral, anti-inflammatory, lowering blood pressure, dispelling wind and clearing heat, calming the liver and improving eyesight. , can be used to treat anemopyretic cold, headache and dizziness, conjunctival congestion and swelling, dim eyesight and other diseases. Different medicinal chrysanthemum varieties have lar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/6895C12Q1/686C12Q2565/125Y02A50/30
Inventor 李敏韩笑杰
Owner ZHEJIANG UNIV OF TECH
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