DNA isolation from dry and fresh samples of polysaccharide-rich plants

• Published in: Plant molecular biology reporter Vol. 20; no. 4; p. 415
• Main Authors: Sharma, Arun Dev, Gill, Prabhjot Kaur, Singh, Prabhjeet
• Format: Journal Article
• Language: English
• Published: New York Springer Nature B.V 01-12-2002
• Subjects: Amplification; Cloning; Contamination; Deoxyribonucleic acid; DNA; Heterogeneity; Metabolites; Plant extracts; Polysaccharides; Saccharides; Seeds; Soybeans; Starches; Wheat
• ISSN: 0735-9640; 1572-9818
• DOI: 10.1007/BF02772129

DNA extraction is difficult in a variety of plants because of the presence of metabolites that interfere with DNA isolation procedures and downstream applications such as DNA restriction, amplification, and cloning. The chemotypic heterogeneity among species may not permit optimal DNA yield with a single protocol; thus, even closely related species may require different isolating protocols. Here we describe a modified procedure based on the hexadecyltrimethylammonium bromide (CTAB) method to isolate DNA from tissues containing high levels of polysaccharides. The procedure is applicable to both dry and fresh tissues and was tested on chickpea seeds, soybean, and wheat leaves. This method solved the problems of DNA degradation, contamination, and low yield due to binding and/or coprecipitation with starches and polysaccharides. The isolated DNA proved amenable to PCR amplification and restriction digestion.