An improved method for the rapid isolation of RNA from Arabidopsis and seeds of other species high in polyphenols and polysaccharides

Abstract

AbstractSeeds are notoriously high in polyphenols and polysaccharides, which reduce RNA quality and yield, and interfere with downstream applications. We present simple modifications to a rapid RNA extraction protocol for use with seeds. The inclusion of polyethylene glycol in place of polyvinylpyrrolidone reduced polyphenol and polysaccharide contamination. In addition, replacing NaCl with KCl improved the RNA yield from Arabidopsis seeds still bound by mucilage. On extraction of Arabidopsis seed recovered from field soils clean RNA pellets with no accompanying gelatinous matrix (polysaccharide) were seen, with A260/230 ratios greater than 1.8 confirming the lack of polysaccharide carry-over. WhenBrassica oleraceaandSinapis arvensisseeds were extracted, 260/230 ratios greater than 1.8 were seen. RNA yields in excess of 10 µg per 100 mg seed suitable for RT-QPCR were obtained.