Journal of Medicinal Plants Research
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Article Number - 15761D914593


Vol.3(1), pp. 016-019 , January 2009

ISSN: 1996-0875



Full Length Research Paper

Optimization of DNA isolation for RAPD-PCR analysis of selected (Echinaceae purpurea L. Moench) medicinal plants of conservation concern from Turkey


Hasibe Cingilli Vural1* and Aslı Dağeri2




1Selcuk University, Department of Biology, Molecular Biology, 42079 Selçuklu, Konya, Turkey.

2Selcuk University, Department of Biology, 42079 Selçuklu, Konya, Turkey.


Email: hcingilli@selcuk.edu.tr






 Accepted: 19 December 2008  Published: 31 January 2009

Copyright © 2009 Author(s) retain the copyright of this article.
This article is published under the terms of the Creative Commons Attribution License 4.0


Genetic analysis of plants relies on high yields of pure DNA samples. Here we present the optimization of DNA isolation protocols and PCR conditions for RAPD analysis of selected medicinal plants of conservation concern from Turkey, containing high levels of polysaccharides, polyphenols and secondary metabolites. These methods involve a modified CTAB extraction employing polyvinyl pyrrolidone while grinding, successive long-term Chloroform-lsoamylalcohol extractions, and EZ1 Nucleic Acid isolation protocols. The yield of DNA ranged from 1 - 2 μg/μl per gram of the leaf tissue and the purity (ratio) was between 1.7 - 1.8 indicating minimal levels of contaminating metabolites. EZ1 nucleic acid isolation technique is ideal for isolation of DNA from different plant species and the DNA isolated was used for randomly amplified polymorphic DNA (RAPD) analysis. RAPD protocol was optimized based on the use of 50 ng of template DNA and annealing temperature of 37°C, resulted optimal amplification. Thus, the results indicate that the optimized protocol for DNA isolation and PCR was amenable to plant species belonging to different genera which is suitable for further work on diversity analysis. Furthermore, here we used suitable DNA isolation protocol for RAPD analysis to study the genetic variation in the future in Echinaceae sp. grown in Turkey.

 

Key words: Echinaceae purpurea, RAPD-PCR, medicinal plant.

 


APA (2009). Optimization of DNA isolation for RAPD-PCR analysis of selected (Echinaceae purpurea L. Moench) medicinal plants of conservation concern from Turkey. Journal of Medicinal Plants Research, 3(1), 016-019.
Chicago Hasibe Cingilli Vural and Aslı Dağeri. "Optimization of DNA isolation for RAPD-PCR analysis of selected (Echinaceae purpurea L. Moench) medicinal plants of conservation concern from Turkey." Journal of Medicinal Plants Research 3, no. 1 (2009): 016-019.
MLA Hasibe Cingilli Vural and Aslı Dağeri. "Optimization of DNA isolation for RAPD-PCR analysis of selected (Echinaceae purpurea L. Moench) medicinal plants of conservation concern from Turkey." Journal of Medicinal Plants Research 3.1 (2009): 016-019.
   
DOI
URL http://academicjournals.org/journal/JMPR/article-abstract/15761D914593

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