Comparative Analysis and Optimization of Different DNA Extraction Protocols in Satureja khuzistanica

Behrooz Doosty, Reza Drikvand, Elham Salahvarzi, Hamzeh Amiri, Javad Hadian

Abstract


Genetic studies of plants are base on high efficiency of purified DNA samples. In this study, we optimized DNA extraction and PCR conditions of Satureja khuzistanica from Iran. The aerial organs of this plant contain high levels of essential oil which makes it difficult to DNA extraction with high quality and thereby intervened with subsequent PCR expansion. Four published DNA extraction protocols include Dellaporta (1983), Doyle and Doyle (1990), Murry and Thompson (1980), Kang and yang (2004) were compared for their capability to produce suitable quality DNA from Satureja khuzistanica. The protocol that provided the foremost DNA quality in the Satureja khuzistanica is selected. In Dellaporta method, no acceptable results were found because SDS buffer extraction was not attached to proteins well. In Doyle and Doyle, the obtained DNA was negligible. In kang and yang method, the quality of extracted DNA was not satisfying. Finally, in modified Murray and Thompson method, the extracted DNA has proper quality. In this method, these factors had the considerable effect on the quality of extracted DNA include: change of incubation time, high NaCl concentration, temperature changes in centrifugation stages, use of proteinase K and TE with high amount of salt and use of plant leaves before flowering stage.

Full Text: PDF DOI: 10.5539/ijb.v4n4p111

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This work is licensed under a Creative Commons Attribution 3.0 License.

International Journal of Biology   ISSN 1916-9671(Print)   ISSN 1916-968X  (Online)

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