Extraction of Genomic DNA from Polysaccharide and Polyphenol Rich Clusterbean (Cyamopsis tetragonoloba (L.) Taub.)

Order of Publishing in Issue: 
5
Volume :9
Issue :1
February, 2015
Page No: 
37-42
Authors: 
S. R. Kalaskar[*], A.S. Shinde, A. H. Rathod., W. A. Sheikh, J. S. Dapke and S. Acharya
Address: 
Department of Plant Molecular Biology and Biotechnology, Centre of Excellence for Research on Pulses, Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar, Gujarat, India-385 506

Clusterbean (2n=14), also known as guar (Cyamopsis tetragonoloba (L.) Taub.) is relatively difficult to work in molecular biology because of the high polysaccharide such as galactomannan content of its endosperm and the high polyphenol content of its leaves. This study aimed to establish a robust genomic-DNA extraction protocol for clusterbean. The efficacy of DNA extraction protocol for a PCR quality genomic DNA extraction was found using DNA fingerprinting. This experiment involves extraction of DNA using a wash buffer containing Tris-HCl, PVP and addition of β- mercaptoethanol separately in each sample and extraction buffer containing CTAB, Tris-HCI, EDTA, NaCl and PVP followed by purification of DNA with RNAase, phenol : chloroform: isoamlyalcohol and finally precipitation of DNA by chilled isopropanol. The protocol is simple and has no special requisites. In terms of quantity (up to1009.5-2119.1 ng/μl) and quality (A260/280 = 1.76 to 1.87) the present method has advantages over many other plant DNA extraction protocols. This protocol can probably be extended also to other leguminous species mainly pulses

Keywords: 
Clusterbean, Guar, Cyamopsis, DNA extraction.
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