100] Isolation and purification of plant nucleic acids from whole tissues and from isolated nuclei

Abstract

This chapter discusses the isolation and purification of plant nucleic acids from whole tissues and isolated nuclei. Three factors are taken into account—(1) Hardness of tissue, (2) pH, and (3) Nucleic acid content. Tissues that are soft pose no problems in mechanical breakage. In a number of plant species, vacuolar pH is low and in such cases, macromolecules may be denatured upon cell injury. Nucleic acids may be isolated and purified from most types of plant tissues, but special procedures are required for tissues that contain a great deal of storage material and/or a great deal of secondary wall growth. A procedure is described in the chapter that separates cytoplasmic and nuclear materials from these interfering components. In the case of plants, tissue bulk is usually a deceptive indicator of the amount of nucleic acid present, and the nucleic acid content should first be determined analytically. The chapter also discusses the preparation of tissues, the fractionation of tissues by nonaqueous procedures and aqueous procedures, preparation of RNA, and preparation of DNA from plant tissues.