Isolating plant genomic DNA without liquid nitrogen

• Published in: Plant molecular biology reporter Vol. 21; no. 1; pp. 43 - 50
• Main Authors: Sharma, R, Mahla, H.R, Mohapatra, T, Bhargava, S.C, Sharma, M.M
• Format: Journal Article
• Language: English
• Published: New York Springer Nature B.V 01.03.2003
• Subjects: Alcohol; Alcohols; Anogeissus acuminata; beans; callus; Chloroform; Cloning; cryopreservation; cumin; Cuminum cyminum; Deoxyribonucleic acid; DNA; DNA fingerprinting; Edetic acid; EDTA (chelating agent); Eruca sativa; Ethylenediaminetetraacetic acids; Fusarium; Fusarium oxysporum; Grinding; isolation; leaves; Liquid nitrogen; Lycium; Lycium barbarum; methodology; millets; Molecular weight; mycelium; Nitrogen; Pennisetum glaucum; Pennisetum typhoideum; Phoenix dactylifera; Plant species; random amplified polymorphic DNA technique; restriction fragment length polymorphism; sequence homology; Tecoma stans; Tecomella undulata; Vigna aconitifolia; Ziziphus mauritiana
• ISSN: 0735-9640; 1572-9818
• DOI: 10.1007/bf02773395

DNA was isolated from leaves of 10 plant species (Cuminum cyminum, Vigna aconitifolia, Pennisetum typhoides, Tecoma stans, Lycium barbarum, Anogeissus acuminata, Tecomella undulata, Zizyphus mauritiana, Phoenix dactylifera, and Eruca sativa) and a fungus (Fusarium oxysporum) using the CTAB method. Three fixing solutions (alcohol, alcohol and chloroform, alcohol and EDTA) were used to produce high molecular weight DNA (>40 kb). DNA quality and quantity was comparable for the 3 fixing solutions and liquid nitrogen grinding. Adding chloroform or EDTA to fixing solutions offered no advantage over absolute alcohol. Isolated DNA was suitable for RAPD analysis, restriction digestion, and cloning. This method does not require liquid nitrogen for fixation, grinding, or storage at -80°C, making it advantagenous over other common protocols.