AGRIVITA, Journal of Agricultural Science
Vol 39, No 3 (2017): OCTOBER

Efficient Strategies for Elimination of Phenolic Compounds During DNA Extraction from Roots of Pistacia vera L.

Saeideh Rajaei (National Institute of Genetic Engineering and Biotechnology, Iran)
Rana Sabagh Farshi (National Institute of Genetic Engineering and Biotechnology, (NIGEB), Iran)
Maryam Moazzam Jazi (National Institute of Genetic Engineering and Biotechnology, (NIGEB), Iran)
Seyed Mahdi Seyedi (National Institute of Genetic Engineering and Biotechnology, (NIGEB), Iran)



Article Info

Publish Date
07 Aug 2017

Abstract

Optimization of DNA extraction protocols for plant tissues and including endophytic microorganisms is a critical step of advanced plant-microbe interaction in agricultural studies. Pistachio (Pistacia vera L.) root tissue contains high levels of polyphenols have been known as major extract contaminants and inhibitors of enzymatic activities during amplification. The present study aimed to develop reliable strategies to purify DNA from Pistachio root samples. Inhibiting substances were removed from DNA through a process including extraction with hot detergent contains SDS-Tris- EDTA, AlNH4(SO4)2.12H2O as chemical coagulating factor and CTAB-NaCl. Following typically organic extraction/alcohol precipitation, denaturing agarose electrophoresis performed to purify probable remain contaminants. The purified DNA was enough free of polyphenols based upon loss of color and spectral quality (260/230>1.6) and efficiently amplified during polymerase chain reaction particularly in the present of GC-clamp primers. This method proved well with detection of Glomus sp. (arbuscular mycorrhiza fungi) associated with Pistacia vera L. using denaturing gradient gel electrophoresis (DGGE).

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Journal Info

Abbrev

AGRIVITA

Publisher

Subject

Agriculture, Biological Sciences & Forestry

Description

AGRIVITA Journal of Agricultural Science is a peer-reviewed, scientific journal published by Faculty of Agriculture Universitas Brawijaya Indonesia in collaboration with Indonesian Agronomy Association (PERAGI). The aims of the journal are to publish and disseminate high quality, original research ...