Abstract
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, andEruca 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.
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Abbreviations
- RAPD:
-
random amplified polymorphic DNA
- RT:
-
room temperature
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Sharma, R., Mahla, H.R., Mohapatra, T. et al. Isolating plant genomic DNA without liquid nitrogen. Plant Mol Biol Rep 21, 43–50 (2003). https://doi.org/10.1007/BF02773395
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DOI: https://doi.org/10.1007/BF02773395