Abstract
The RNA fraction was extracted from leaves of Eucalyptus, six other woody plants and four herbaceous plants with an extraction buffer containing sodium isoascorbate at a concentration of 500 mM. This method consisted of one or two chloroform extractions, one acid guanidium-phenol-chloroform extraction and isopropanol precipitation alone. The yields of the RNA fractions and the ratios of absorbance at 260 nm/280 nm and 260 nm/230 nm were 246-1750 μg-1 fresh weight, 1.94-2.03 and 2.07-2.50, respectively. When the RNA fractions obtained with this method were subjected to agarose gel electrophoresis, intact rRNA bands were detected. When the reverse transcript-PCR reaction was performed using these RNA fractions and de-generated primers for NADP-specific isocitrate dehydrogenase, an amplified fragment at ≈1.1 kb was obtained as expected. These results indicate that our new method achieves a simple and rapid preparation of high quality RNA from leaves of Eucalyptus and other plant species.
