Extracting high-yield, high-quality DNA from plant samples is challenging due to the presence of the cell wall, pigments, and some secondary metabolites. The main CTAB method, two of its modified protocols (beta-mercaptoethanol or ammonium acetate were eliminated), the modified Murray and Thompson method, and the Gene All kit were statistically compared based on the quantity and quality of the total DNA (tDNA) extracted from fresh and dried leaves of three medicinal herbs P. harmala, T. ramosissima, and P. reptans. The suitability of the tDNAs for molecular studies was evaluated by polymerase chain reaction (PCR) of the fragments of the internal transcribed spacer (ITS) in nuclear DNA and the trnL-F region in chloroplast DNA. Some significant differences were found between the tDNAs extracted by five extraction methods. With the exception of P. harmala, where the PCR of both the ITS fragments and the trnL-F region worked successfully in all DNA samples, but only the ITS fragments, not the chloroplast trnL-F region, were amplified in the DNA samples of T. ramosissima and P. reptans. The chloroplast trnL-F region was amplified only in DNA samples extracted from fresh and dried leaves of the three studied herbs using the commercial kit. Gene All kit, the main CTAB method, and its modified protocols were the less time-consuming protocols that yielded DNA suitable for downstream PCR vis-a-vis the modified Murray and Thompson method.
Purpose Traditional medicine and herbal therapy, as a health-care approach of traditional medicine, are receiving increased attention from both the public and scientific society. The number of articles presented on traditional medicine and its applications is increasing rapidly. However, compared with extensive ongoing herbal research, achievements are not remarkable. The objective of the present article is to highlight some existing research gaps, which bridging the gaps will increase the quality and validity of the results of herbal research. Methods Iranian and non-Iranian articles on Allium sativum L. were reviewed for compliance with standards and requirements for herbal research. Results Botanical nomenclature, the processing and the extraction procedures, and the history of traditional medicine were the most important gaps in the herbal research. Conclusions Traditional and herbal medicines are rich and promising sources for drug development. Researchers and authors should consider that the familiarity with the history, characteristics, and potentials of traditional medicine systems is an important factor, accelerating the process of natural-based drug development. Moreover, essential standards with a high impact on the validity and reliability of the results of herbal research must not be ignored. Also, numerous fields remain to be considered by scientists and researchers, including the status of medicinal herbs, toxicological assessments, biological and pharmacological mechanisms of disease, and research questions.
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