Easy DNA Extraction Protocol for Ricinus communis Seeds | InformativeBD

Ajmal Iqbal,  Waqar Ahmad,  Asaf Khan,  Murad Khan, and Mohammad Nisar, from the institute of Pakistan,. wrote a Research article about, Easy DNA Extraction Protocol for Ricinus communis Seeds. Entitled, User friendly DNA isolation protocol optimized for Ricinus communis L. seeds. This research paper published by the International Journal of Biosciences | IJB.  an open access scholarly research journal Biosciences. under the affiliation of the International Network For Natural Sciences| INNSpub. an open access multidisciplinary research journal publisher.

Abstract

A protocol was developed to isolate high quality genomic DNA from the seeds of Ricinus communis L. (caster) without using liquid nitrogen.The DNA extraction buffer used in this novel protocol constitutes SDS (1%), Tris (1.21%), NaCl (0.58%), EDTA (0.32%), 0.12% and β–Mercaptoethanol witha pH 8.5. In the protocol 0.09g of crushed seeds of caster bean, 600ul of DNA extraction buffer and and 500ul of phenol: chloroform: iso-amylalcohol with a ratio of 25:24:1 were used. The isolated DNA was amplified in Polymerase Chain Reaction using RAPD and SSR primer sets. The primer set successfully amplified the isolated DNA. Hence, the protocol is recommended as a user friendly novel protocol for DNA isolation from the castor beans seeds.

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Introduction

Ricinus communis L., commonly known as caster bean, is a member of the spurge family,Euphorbiaceae (Smith 1986). Ricinus is indigenous to tropical Asia and Africa, but today it is cultivated for seed oil throughout the tropical and subtropical regions of the world (Seo, 2011). The annual production of caster bean throughout the world is 460,000 tones. In Pakistan it is grown on 3204 ha and its annual production is 2089 tones, the average seed yield of caster bean in Pakistan is 652 kg ha-1 . This seed yield of Pakistan is very low (Anonymous, 2006). Oil derived from the seeds of caster has several potentials in many industries like medicine, and cosmetics (Akande, 2012). Castor bean seed oil is highly valued in several sectors of the chemical industry and is considered a bioenergy and phytoremediation resource in the subtropics (Baldanzi et al. 2005). Castor plant has a great drought tolerance due to deep root system with the ability to explore the deeper layers of soil, which helps increase aeration, water retention and distribution in soil (Embrapa et al., 2006).

The Preliminary Phytochemical study of Ricinus communis revealed the presence of steroids, saponins, alkaloids, flavonoids, and glycosides (Kang, et al., 1985). The leaves of the Ricinus communis contain flavonoids, tannins and phenol (Yadav RNS & Agarwala M. 2011; Ilavarasan et al., 2006).

The isolation of high-quality DNA is prerequisite for any molecular biology work because contaminants such as proteins, polyphenols and polysaccharides may interfere with enzymes, such as endonuclease (in blotting techniques) and Taq polymerase in Polymerase Chain Reaction (Ausubel et al., 1994). The phenols covalently bind to proteins and DNA, giving the DNA a brown colour and making it useless for most research applications (Katterman & Shattuck, 1983; Guillemaut & Drouard, 1992, Aljanabi et al., 1999). Polyphenol contamination of DNA makes it resistant to restriction enzymes (Katterman & Shattuck, 1983).

Several methods for extracting DNA for different plant are available (Doyle and Doyle et al., 1990;

Khanuja et al., 1999; Kumar et al., 2003 Islam et al., 2013). In Pakistan no work has been done on the extraction of DNA from Ricinus species. Therefore the aim of the present study is to develop simple DNA extraction protocol from the seeds of six different varieties of Ricinus communis without liquid nitrogen and for further genome characterization using RAPD and SSR primers.

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