99] The isolation of nucleic acids from bacterial spores

Abstract

Publisher Summary This chapter discusses the isolation and purification of plant nucleic acids from whole tissues and from isolated nuclei. Tissues that are soft pose no problems in mechanical breakage (for example, meristematic regions). Specific methods for some types of tissue are provided in the chapter, but it is emphasized that plant cell walls are formidable barriers to the diffusion of large molecules. Incomplete cell breakage may result not only in low yields of nucleic acid but also what is more important; preferential breakage of young cells results in selective extractions. In a number of plant species, vacuolar pH is low and in such cases, macromolecules may be denatured upon cell injury. Vacuolar membranes are likely to be disrupted before cells are broken and Tris or phosphate buffers may not penetrate rapidly enough to offset vacuolar acidity. If preprocessing with organic solvents is not desired, prior infiltration of tissue pieces with dilute ammonium hydroxide (e.g., 0.001 M) should be tried. Nucleic acids may be isolated and purified from most types of plant tissues.