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Zhebaizhu pcr identification kit and identification method

A kit and Atractylodes Rhizoma technology, applied in the field of DNA molecular identification, to achieve the effects of high sensitivity, accurate detection, and promoting the modernization and development of traditional Chinese medicine

Active Publication Date: 2017-01-18
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the identification of Atractylodes Rhizoma Atractylodes Rhizoma by PCR detection method

Method used

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  • Zhebaizhu pcr identification kit and identification method
  • Zhebaizhu pcr identification kit and identification method

Examples

Experimental program
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Effect test

Embodiment 1

[0030] 1. Acquisition of PCR primers for key reagents of Zhebaizhu PCR identification kit

[0031] Genomic DNA was extracted from Zhejiang Baizhu, Hebei Xiaobaizhu, Hunan Pingzhu and Anhui Huishu using the Plant Genomic DNA Rapid Extraction Kit (Shanghai Sangon Bioengineering Co., Ltd.). Using the RAPD amplification method to perform RAPD analysis on the four Atractylodes macrocephala, a specific amplified band of Atractylodes atractylodes was screened, and this specific amplified band only appeared in the variety of Atractylodes atractylodes, but not in other varieties of Atractylodes atractylodes. The DNA molecule in the band is the specific DNA molecular marker of Atractylodes macrocephala. The specific DNA molecular marker of Atractylodes macrocephala was recovered from the agarose gel, cloned and sequenced. The sequencing result is shown in SEQ ID NO.3 and named ZBZ647.

[0032] According to the ZBZ647 sequence, design a pair of specific primers ZBZ1 and ZBZ2, the primer...

Embodiment 2

[0048] 1. Study on the accuracy of detection method of Zhejiang Atractylodes macrocephala PCR identification kit

[0049] Genomic DNA was extracted from 14 species of plants: Zhejiang Atractylodes, Hebei Xiaobaizhu, Hunan Pingzhu, Anhui Huishu, Cangzhu, Guantract, Hangbaiju, Wildchrysanthemum, Polygonatum, Dandelion, Sunflower, Zhejiang Fritillaria, Hang Maidong and Motherwort. Using the genomic DNA of the above 14 varieties as a template, PCR amplification was performed using specific primers "ZBZ1 / ZBZ2". 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 PCR of Atractylodes atractylodes. The accuracy and reliability of the detection method, the results see figure 1 As shown, lanes 1 to 14 are Zhejiang Atractylodes, Hebei Xiaobaizhu, Hunan Pingzhu, Anhui Huishu, Cangzhu, Guantract, Hangbaiju, Wild Chrysanthemum, Polygonatum, Dandelion, Sunflower, Zhejiang Fritillaria, ...

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Abstract

The invention discloses a rhizoma atractylodis macrocephalae PCR identification kit and an identification method thereof; specific primers of the kit comprise ZBZ1 and ZBZ2, ZBZ1 is 5'-CAAGTTGGATACACTCCTGG-3, and ZBZ2 is 5'-CTACTACCATCGCAAACTCT-3'; the invention provides the method for fast and accurate identification of rhizoma atractylodis macrocephalae; the method can widely applied in variety identification of various links of rhizoma atractylodis macrocephalae traditional Chinese medicine seed selection and seed breeding, planting, harvesting and storage, sale and clinical application, and has great significances in identifying the authenticity of the traditional Chinese medicine, combating counterfeits, strengthening the quality supervision of the traditional Chinese medicine, and promoting the modernization development of the traditional Chinese medicine.

Description

[0001] (1) Technical field [0002] 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 for traditional Chinese medicine Atractylodes macrocephala. [0003] (2) Background technology [0004] Atractylodes macrocephala is the dried rhizome of Atractylodes macrocephala Koidz., which is widely used clinically. , the effect of antiperspirant and antiabortive. Zhejiang is the main production area of ​​Atractylodes atractylodes, with a cultivation history of hundreds of years. The varieties produced in Pan'an, Xinchang, Shengxian, Dongyang and other places in Zhejiang Province are called "Zhebaizhu" (Rhizoma Atractylodis Macrocephalae), which is one of the famous authentic medicinal materials "Zhebawei". It is characterized by its relatively plump shape, " The "operated legs" are thick and short, and the "cloud head" is swollen. [0005] In recent years, due to the con...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2531/113
Inventor 李敏陈强茅学群黄龙妹
Owner ZHEJIANG UNIV OF TECH
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