title: Isolation of nucleic acids from plants by differential solvent precipitation.
abstract: 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.
authors.affiliation: Department of Plant Physiology, Institute of Horticultural Research, West Sussex, England.
authors.forename: K
authors.initials: K
authors.lastname: Manning
mesh_terms: D000433:1-Propanol; D002021:Buffers; D002241:Carbohydrates / Q000032:analysis; D011232:Chemical Precipitation; D002712:Chlorides; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005026:Ethylene Glycols; D005419:Flavonoids; D008094:Lithium; D018021:Lithium Chloride; D009696:Nucleic Acids / Q000302:isolation & purification*; D010636:Phenols / Q000032:analysis; D010779:Photofluorography; D010944:Plants / Q000235:genetics*; D011108:Polymers / Q000032:analysis; D059808:Polyphenols; D014176:Protein Biosynthesis / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012997:Solvents
chemical_list: D002021:Buffers; D002241:Carbohydrates; D002712:Chlorides; D005026:Ethylene Glycols; D005419:Flavonoids; D009696:Nucleic Acids; D010636:Phenols; D011108:Polymers; D059808:Polyphenols; D012997:Solvents; D012313:RNA; D000433:1-Propanol; D008094:Lithium; D018021:Lithium Chloride; C017096:n-butoxyethanol