Improved method for the isolation of RNA from plant tissues.
A fast and efficient method for the isolation of RNA from plant tissues is described. Tuber tissue is homogenized in a guanidine hydrochloride-containing buffer followed by direct extraction with phenol/chloroform. The RNA is precipitated from the aqueous phase, washed with 3 M sodium acetate and 70% ethanol, and finally dissolved in water. The yield of RNA is up to 500 micrograms/g of tissue and several tests indicate intact and nondegraded RNA. This method can be adapted to a small-scale version by the use of 1.5-ml tubes, allowing rapid isolation of RNA from a larger number of samples. Finally, this method is of particular use for isolating RNA from tissues with a high polysaccharide and nuclease content such as wounded potato tubers.
D002725:Chloroform; D004587:Electrophoresis, Agar Gel; D000431:Ethanol; D019791:Guanidine; D006146:Guanidines; D009693:Nucleic Acid Hybridization; D000439:Pentanols; D019800:Phenol; D010636:Phenols; D010947:Plants, Toxic; D012313:RNA / Q000302:isolation & purification*; D011198:Solanum tuberosum / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics*
D006146:Guanidines; D000439:Pentanols; D010636:Phenols; D019800:Phenol; D000431:Ethanol; D012313:RNA; D002725:Chloroform; C029683:isopentyl alcohol; D019791:Guanidine