A method for high-quality RNA extraction from tall fescue

Abstract

The isolation of high-quality RNA was a precondition in molecular biology research of tall fescue. Two common approaches were adopted for the total RNA extraction by using leaves of tall fescue as the material in this experiment in order to seek the optimized total RNA extraction method of tall fescue, as well as the improvement of the extraction method used by the predecessors. The results showed that the Trizol method cost much and was not suitable for the large quantity of plant tissues extraction. The traditional isothiocyanate method resulted in protein contamination and RNA degradation. By employing the improved isothiocyanate method, we found that there were three bright bands in agarose gel electrophoresis (28S rRNA, 18S rRNA and 5S rRNA). The band of 28S rRNA was brighter than that of 18S rRNA, and the value of OD₂₆₀/OD₂₈₀ was 1.8 to 2.0. Clear bands and high polymorphisms were obtained by cDNA-AFLP analysis. These results indicated that the RNA isolated by the improved isothiocyanate method had a good integrity and high purity, which could be used for the later molecular researches.

Key words: Tall fescue, RNA extraction, improved isothiocyanate method.