Isolation of functional total RNA from Argemone mexicana tissues

Abstract

RNA extraction from recalcitrant plant tissues is frequently complicated by the presence of secondary metabolites, polysaccharides and polyphenols. These compounds may co precipitate with RNA, often rendering it unsuitable for either cDNA synthesis or hybridization in northern blot analyses and therefore, interfering with the gene analysis expression in such tissues. We have developed an efficient RNA extraction method from A. mexicana tissues. The procedure includes the use of polyvinylpolypyrrolidone (PVPP), to remove secondary metabolites, proteins and polyphenols, and a two-step precipitation with LiCl, to eliminate polysaccharides, and thus increasing RNA yield. The quality of the resulting RNA was evaluated spectrophotometrically and by agarose gel electrophoresis. Moreover, the RNA obtained by this method, could be used directly for both RT-PCR and northern blot analysis, without any further purification.