Comparison study of three methods for genomic DNA extraction from fresh and herbarium leaf specimens of Achillea wilhelmsii C. Koch

Azra Saboora | Tayebeh Radjabian | Maryam Amiri Rad | Ezat Asgarani

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URL :   http://research.shahed.ac.ir/WSR/WebPages/Report/PaperView.aspx?PaperID=116875
Date :  2019/03/19
Publish in :    يافته هاي نوين در علوم زيستي

Link :  https://nbr.khu.ac.ir/article-1-2974-fa.html
Keywords :Achillea wilhelmsii, genomic DNA extraction, herbarium samples, magnetic nanoparticle, polyphenols

Abstract :

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DNA extraction from plant tissues is often problematic. For example, unsuccessful removal of secondary metabolites, such as phenolic compounds, during the extraction of the DNA of aromatic and medicinal plants results in errors in the results of molecular experiments. Achillea wilhelmsii is a medicinal plant which belongs to Asteraceae family and is native to Iran, there is little information about its genomic data. Therefore, the optimization of DNA extraction methods to obtain yield with higher quality and quantity is necessary. In this study, two traditional DNA extraction methods (using fresh and herbarium leaf samples) and a commercial DNA kit (MAGNANTM) in Yarrow were compared. Results showed that DNA extracted from fresh leaves of yarrow by the application of Khanuja et al. (1999) method was better than other methods mentioned in this research, because of increased amount of extracted DNA and reduced amount of harmful compounds such as RNA, polysaccharides, protein and secondary metabolites. The evaluation of the quality of extracted DNA from herbarium specimens showed that the quality of this extract was low on agarose gel because of the formation of smear and broken molecules, in spite of the high yielding DNA (10-50 fold ratio to the other methods). This can be considered to be a limitation of the extract in molecular experiments. The procedure of MAGNANTM DNA kit proved to be appropriate for reducing the time and cost of extraction as well as its low contamination of protein and RNA. However, this process still needs some modifications for yarrow to increase the amount of extracted DNA.