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Hebei Baizhu pcr identification kit and identification method

The technology of a kit and Atractylodes macrocephala is applied in the field of DNA molecular identification to achieve the effect of simple operation, high sensitivity and accurate identification

Active Publication Date: 2017-04-12
邳州市润宏实业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The methods for accurate identification of Atractylodes atractylodes reported in the literature are all traditional Chinese medicine identification methods, and the identification of Atractylodes Hebei by PCR detection method has not been reported yet.

Method used

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  • Hebei Baizhu pcr identification kit and identification method
  • Hebei Baizhu pcr identification kit and identification method
  • Hebei Baizhu pcr identification kit and identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1. Acquisition of PCR primers for key reagents of Hebei Atractylodes macrocephala PCR detection method

[0032] Genomic DNA was extracted from Zhejiang Atractylodes Rhizoma, Hebei Atractylodes Rhizoma Atractylodes Rhizome, Hunan Pingzhu and Anhui Atractylodes Rhizoma using the Rapid Genomic DNA Extraction Kit. Using the RAPD amplification method to perform RAPD analysis on the four Atractylodes macrocephala, a specific amplified band of Hebei Atractylodes macrocephala was screened, and this specific amplification band only appeared in Hebei Atractylodes atractylodes species, but not in other Atractylodes macrocephala species. The DNA molecule in the band is the specific DNA molecular marker of Hebei Atractylodes macrocephala. The Hebei Baizhu 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 HBZ835.

[0033] According to the sequence of HBZ835, design a pair of specific primer...

Embodiment 2

[0049] 1. Study on the Accuracy of Hebei Baizhu PCR Detection Method

[0050] Genomic DNA was extracted from 14 plants including Zhejiang Atractylodes Atractylodes, Hebei Atractylodes Atractylodes, Hunan Pingzhu, Anhui Huishu, Cangzhu, Guantract, Hangbaiju, Wild Chrysanthemum, Polygonatum, Dandelion, Sunflower, Fritillaria fritillaria, Ophiopogon japonicus and Motherwort. Using the genomic DNA of the above 14 varieties as a template, PCR amplification was performed using specific primers "HBZ1 / HBZ2". 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 Hebei Baizhu PCR. The accuracy and reliability of the detection method, the results see figure 1 As shown, the lanes 1 to 14 are Hebei Atractylodes, Zhejiang Atractylodes, Hunan Pingzhu, Anhui Huishu, Atractylodes atractylodes, Atractylodes officinalis, Hangbaiju, wild chrysanthemum, dandelion, sunflower, Polygonatum, Zhejiang ...

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Abstract

The invention discloses a PCR (polymerase chain reaction) identification kit and identification method for largehead atractylode in Hebei. Specific primers in the kit are HBZ1 and HBZ2, wherein the sequence of the HBZ1 is 5'-GGACATGACAGAAGTATG-3', and the sequence of the HBZ2 is 5'-TGCTGAGAGAAGCAAATC-3'. The invention provides a method for rapidly and accurately identifying largehead atractylode in Hebei, which can be widely applied to the variety identification in each link of seed selecting and breeding, planting, harvesting, storage, selling and clinical applications of a traditional Chinese medicine largehead atractylode in Hebei, and has great significance in the aspects of identifying the authenticity of traditional Chinese medicines, cracking down counterfeit drugs, strengthening the quality supervision of Chinese traditional medicines, and promoting the modernized development of traditional Chinese medicines.

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 of a traditional Chinese medicine Hebei Atractylodes macrocephala. [0003] (2) Background technology [0004] Atractylodes macrocephala is the dried rhizome of Atractylodes macrocephala Koidz., a plant of the Compositae family. It has various medicinal functions such as invigorating the spleen and replenishing qi, drying dampness and promoting diuresis, antiperspirant, and preventing miscarriage. It is widely used in clinical practice. There are many varieties of Atractylodes macrocephala, the main varieties include Hebei Atractylodes, Zhejiang Atractylodes, Hunan Pingshu, and Anhui Huishu, which are produced in Hebei, Zhejiang, Hunan and Anhui respectively. Among them, Hebei Baizhu, also known as "Xiaobaizhu", has the highest yield, and its medicinal ingredients are somewhat di...

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 邳州市润宏实业有限公司
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