Comparative Different DNA Isolation Protocols from Ziziphus spina-christi (L.) Leaves through RAPD and ISSR Markers

Fatemeh Bina, Zabihollah Zamani, Vahideh Nazeri, Daryush Talei

Abstract


Genomic analysis of plants relies on high quantity and quality of pure DNA. Extraction and purification of DNA from woody and medicinal plants, such as fruit trees present a great challenge due to accumulation of a large amount of co-purify with DNA, including polysaccharides, polyphenols and proteins. Therefore, it is necessary to optimize the extraction protocols to reduce these compounds to the lowest level. A study was conducted to compare six DNA extraction and precipitation methods for genomic analysis in Ziziphus spina-christi (L.) plant tissues. The results showed significant differences in DNA contents among the six methods. Quantity and quality of extracted genomic DNAs were compared by employing the spectrophotometer, Nano-Drop, agarose gel electrophoresis, digestion by restriction enzymes and polymerase chain reaction (PCR) methods and molecular marker such as RAPD and ISSR. The method of Vroh Bi et al., provided the best results (208.89 ng/μL) in terms of quantity and quality of DNA, and Doyle and Doyle method as second method for leaves sample were chosen. According to the results, the method of Bi et al. is recommended for DNA extraction from plant tissues having high level of polysaccharides and phenol compounds.


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DOI: http://dx.doi.org/10.5539/jas.v8n6p49

Journal of Agricultural Science ISSN 1916-9752 (Print) ISSN 1916-9760 (Online)

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