Isolation of nucleic acids from plants by differential solvent precipitation.
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.
Manning
K
K
Department of Plant Physiology, Institute of Horticultural Research, West Sussex, England.
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
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