lithium-chloride and n-butoxyethanol

The purification of nucleic acids from plant tissue is often made difficult by the presence of contaminating carbohydrate polymers and polyphenols. A procedure for the simultaneous isolation of DNA and translatable RNA from plants is described. The method removes most of the polysaccharides and polyphenols extracted with nucleic acids in a single step by taking advantage of differences in solubility of these compounds in the solvent 2-butoxyethanol. Stepwise addition of 2-butoxyethanol to phenol extracts of specific ionic strength precipitates nucleic acids largely free of contaminants. Subsequent separation of RNA from DNA by precipitation with LiCl was optimised to give a high recovery of translationally active RNA. Successful isolation of nucleic acids from strawberry (Fragaria X ananassa) receptacle, a particularly recalcitrant tissue, and from a range of tissues of other plant species demonstrates the general applicability of the method.

Topics: 1-Propanol; Buffers; Carbohydrates; Chemical Precipitation; Chlorides; Electrophoresis, Agar Gel; Electrophoresis, Polyacrylamide Gel; Ethylene Glycols; Flavonoids; Lithium; Lithium Chloride; Nucleic Acids; Phenols; Photofluorography; Plants; Polymers; Polyphenols; Protein Biosynthesis; RNA; Solvents