A modified genomic DNA extraction method from leaves of sunflower for PCR based analyzes

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Research Paper 01/12/2015
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A modified genomic DNA extraction method from leaves of sunflower for PCR based analyzes

Hossein Zeinalzadehtabrizi, Arash Hosseinpour, Murat Aydin, Kamil Haliloglu
J. Biodiv. & Environ. Sci. 7(6), 222-225, December 2015.
Copyright Statement: Copyright 2015; The Author(s).
License: CC BY-NC 4.0

Abstract

Sunflower genomic DNA extraction for PCR (Polymerase Chain Reaction) analyses and similar molecular technics is a challenging issue because of high concentrations of secondary metabolites, i.e. polyphenolics, polysaccharides, tannins and other contaminants. Our modified (Cetyl Trimethyl Ammonium Bromide) CTAB protocol can be performed approximately in two hours and suitable for PCR analyses. It needed small amounts of plant leaves and good quality of sunflower genomic DNA can be isolated. Quantity and purity of extracted DNA were checked by spectrophotometry and agarose gel electrophoresis. This modified method takes less time, reliable and gives good quality of DNA even without using RNase can be used in various marker systems such as (Single Sequence Repeat) SSR and (Random Amplified Polymorphic DNA) RAPD.

Amani J, Kazemi R, Abbasi AR, Salmanian AH. 2011. A simple and rapid leaf genomic DNA extraction method for polymerase chain reaction analysis. Iran J Biotechnol. 9, 69-71.

Encheva J, Köhler H, Christov M, Friedt W. 2005. Intergeneric hybrids between cultivated sunflower (Helianthus annuus L.) and Verbesina helianthoides (genus Verbesina): RAPD analysis. Heila 28(42), 37-44.

Horne EC, Kumpatla SP, Patterson KA, Gupta M, Thompson SA. 2004. Improved high-throughput sunflower and cotton genomic DNA extraction and PCR fidelity. Plant Molecular Biology Reporter 22(1), 83-84.

Iqbal M, Sadaqat H, Khan I. 2008. Estimation of genetic diversity among sunflower genotypes through random amplified polymorphic DNA analysis. Genet. Mol. Res. 7(Suppl 4), 1408-1413.

Isaacs MS, Manivannan N, Muralidharan V. 2003. Genetic diversity analysis using RAPD marker in inbred lines of sunflower Helianthus annuus L. Helia 26(39), 59-65.

Li J, Yang J, Chen D, Zhang X, Tang Z. 2007. An optimized mini-preparation method to obtain high-quality genomic DNA from mature leaves of sunflower. Genet. Mol. Res 6(4), 1064-1071.

Zhizhong X. 2008. DNA extraction from oil sunflower seeds and the optimization of its RAPD reaction system. Journal of Anhui Agricultural Sciences 36(6), 2259.

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