Preparation of Genomic DNA from Plant Cells

Abstract

The isolation of undegraded DNA from plants is notoriously difficult, presenting problems that are not, in general, encountered in purification of DNA from animal or bacterial cells. Plant cells have a very tough cell wall, the breakage of which requires the application of vigorous physical force that can shear large DNA molecules. Also, plant cells accumulate large amounts of sencondarymetabolities in their vacuoles that either co-purify with nucleic acids (e.g., polysaccharides) or interact with DNA (polyphenols, oxalic acid, etc.) rendering the final product unusable. Finally, plant cells contain large amounts of very active nucleic acid degrading enzymes, making the time between cell rupture and inactivation of these nucleases a critical factor.