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

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Abstract:

DNA extraction from plant tissues often causes most problems. For example, unsuccessful removal secondary metabolites during extraction, such as phenolic compounds in aromatic and medicinal plants, cause to some mistakes in result of molecular experiments by using of the extracted DNA. Achillea wilhelmsii is a medicinal plant belong to Asteraceae family and native to Iran, there is little information about genomic data in this plant. Therefore, optimizing of the DNA extraction methods for obtaining suitable quality and quantity yield is necessary. In this study, two traditional DNA extraction methods (using fresh and herbarium leaf samples) and commercial DNA kit MAGNANTM in Yarrow have been compared. Results showed that DNA extracted from fresh leaves of yarrow according to Khanuja et al. (1999) was better than the other methods which mentioned in this research, because of the increased amount of extracted DNA and reduced harmful compounds such as RNA, polysaccharides, protein and secondary metabolites. To evaluate the quality of extracted DNA from herbarium specimens showed that, in spite of the high yielding DNA (10-50 fold ratio to the other methods), quality of this extract was low on agarose gel because of smear and broken down the molecules. It can be considered as a limitation of the extract in molecular experiments. Procedure of MAGNANTM DNA kit was appropriate for reducing time and cost of the extraction as well as low contamination to protein and RNA, but the process needs some modifications for yarrow to increase the amount of extracted DNA.

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Journal title

volume 5  issue 4

pages  458- 465

publication date 2019-03

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