Study on the Ethnic Medicine Column

RFLP-HRM identification method of Arnebiae Radix based on DNA barcoding and high resolution melting*

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  • 1. National Institutes for Food and Drug Control, Beijing 102629, China;
    2. China Pharmaceutical University, Nanjing 210009, China;
    3. Songyuan Institute for Food and Drug Control, Songyuan 138000, China

Received date: 2022-01-17

  Online published: 2024-06-24

Abstract

Objective: To establish an identification method of restriction fragment length polymorphism-high resolution melting(RFLP-HRM) for Arnebiae Radix from medicine market based on DNA barcoding and high resolution melting(HRM). Methods: Internal transcribed spacer Ⅱ of nuclear ribosomal DNA (ITS2) sequences of Arnebiae Radix were amplified with polymerase chain reaction(PCR). MEGA 6.06 software was used to perform NJ tree cluster analysis and genetic distance analysis on the ITS2 sequence of Arnebiae Radix. Original plant confirmation of non-pharmacopoeial Arnebiae Radix from the market was performed. The primers were disigned using the software of Primer Premier 5 based on ITS2 sequence. The best one from 4 designed primers was screened through the HRM experiment. The identification method of RFLP-HRM for Arnebiae Radix was established. Results: The neighbour joining clustering analysis and genetic distance analysis of ITS2 sequences could effectively distinguish among the different species for this study. The HRM experiment confirmed Zcf-1 was the best primer. The RFLP-HRM method to effectively identify Arnebiae Radix from medicine market was established. Conclusion: The methods of DNA Barcoding and HRM both can effectively identify the Arnebiae Radix in this study, and the results are consistent and mutual authentication. The RFLP-HRM rapid identification method of Arnebiae Radix from medicine market has been established, which provides the reference basis for other traditional medicines.

Cite this article

LIU Jie, FANG Wen-liang, TANG Zhe, LIAN Chao-jie, DAI Sheng-yun, GUO Li-nong, ZHENG Jian, QIAO Fei, MA Shuang-cheng, LI Yun-zheng . RFLP-HRM identification method of Arnebiae Radix based on DNA barcoding and high resolution melting*[J]. Chinese Journal of Pharmaceutical Analysis, 2022 , 42(8) : 1354 -1362 . DOI: 10.16155/j.0254-1793.2022.08.08

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