PJB-2019-1050
Comparative study of genomic DNA extraction protocols of Tamarix L. species from Iran
Shahram Mehri
Abstract
ABSTRACT The genus Tamarix L. with about 54 species that mainly grow in saline areas of deserts and semi-deserts, is chemically characterized by the presence of tannins, flavonoids, anthocyanins and essential oils which interfere with the extraction of pure genomic DNA. It is necessary to optimize the extraction protocols to reduce the effects of the presence of these compounds to the lowest level. The present study compares the plant genomic DNA extraction Kit (DNP TM Kit ), CTAB DNA extraction method by Murray and Thompson, Sahu et al., Nalini et al. and Bi et al., from the extracting DNA point of view Tamarix species. The results showed significant differences in DNA contents between the five methods. Quantity and quality of extracted genomic DNAs were compared by employing the spectrophotometer, Nano-Drop, agarose gel electrophoresis, and polymerase chain reaction (PCR) methods and molecular marker such as ITS and ISSR. The method of Nalini et al., provided the best results (207 ng/μL) in terms of quantity and quality of DNA, therefore, this method was taken and optimized for DNA extraction. Our results proposed that this method could be effective for plants with same polysaccharides, proteins and polyphenols components. The advantage of this method is simple and fast as it does not involve time consuming steps such as incubation at higher temperatures, does not require expensive chemicals such as proteinase K, liquid nitrogen etc., and no special equipment is needed. The success of this method in obtaining high-quality genomic DNA has been demonstrated in the Tamarix species group and the reliability of this method has been discussed.
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