Isolation of nucleic acids from plants by differential solvent precipitation

Anal Biochem. 1991 May 15;195(1):45-50. doi: 10.1016/0003-2697(91)90292-2.

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.

Publication types

  • Comparative Study

MeSH terms

  • 1-Propanol
  • Buffers
  • Carbohydrates / analysis
  • Chemical Precipitation
  • Chlorides
  • Electrophoresis, Agar Gel
  • Electrophoresis, Polyacrylamide Gel
  • Ethylene Glycols*
  • Flavonoids*
  • Lithium
  • Lithium Chloride
  • Nucleic Acids / isolation & purification*
  • Phenols / analysis
  • Photofluorography
  • Plants / genetics*
  • Polymers / analysis
  • Polyphenols
  • Protein Biosynthesis / genetics
  • RNA / genetics
  • RNA / isolation & purification
  • Solvents*

Substances

  • Buffers
  • Carbohydrates
  • Chlorides
  • Ethylene Glycols
  • Flavonoids
  • Nucleic Acids
  • Phenols
  • Polymers
  • Polyphenols
  • Solvents
  • RNA
  • 1-Propanol
  • Lithium
  • Lithium Chloride
  • n-butoxyethanol