Extraction of High-quality Intact DNA from Okra Leaves Despite Their High  

Nisar Ahmed1 , Sehar Nawaz1 , Ahsan Iqbal1 , Muhammad Mubin1 , Aaish Butt1 , David Anderson Lightfoot2 , Masahiko Maekawa3
1 Centre of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture, Faisalabad, 38000, Pakistan
2 Department of Plant, Soil and Agricultural Systems, Mailcode 441205 Lincoln Drive, Carbondale IL62901, USA
3 Institute of Plant Science and Resources, Okayama University, 2-20-1, Chuo, Kurashiki, 710-0046, Japan
Author    Correspondence author
Bioscience Methods, 2013, Vol. 4, No. 4   doi: 10.5376/bm.2013.04.0004
Received: 29 Apr., 2013    Accepted: 28 May, 2013    Published: 04 Jun., 2013
© 2013 BioPublisher Publishing Platform
This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract

The presence of mucilaginous acidic polysaccharides in okra leaves interferes with the extraction of high-quality intact DNA for later restriction digestion and PCR amplification. We have developed a simple, efficient and reliable protocol to extract high-quality, intact DNA from the highly mucilaginous leaves of okra. The isolated DNA was free of contaminating agents like polysaccharides, protein and polyphenols and (A/260: A/280) ratio of 1.6-2.1 indicate minimal presence of other contaminating metabolites. The extracted DNA was digested with restriction enzyme AvaII and analyzed by PCR using random amplified polymorphic DNA primers. The extraction protocol is simple and does not require liquid nitrogen. The yield and quality of the resulting DNA are satisfactory, and the protocol can be scaled-up.

Keywords
Okra; DNA extraction; Mucilaginous acidic polysaccharides; SDS
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pornliz suckporn sex videos bbw mom xxx big fucking arabin porn videos teen gril sex video riding hard cock woman hard vagina . Nisar Ahmed
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