Separation of low molecular weight RNA species by high-speed gel filtration

By employing a high-speed gel filtration method using a column of silica-based aqueous gel, TSK-GEL G 300 SW, the three low molecular weight RNA species, 5.8S and 5S rRNA, and tRNA, were clearly separated from each other and from high molecular weight RNA species in 200 mM sodium phosphate buffer, pH 7.0, containing 0.1% SDS. Excellent reproducibility and resolution were obtained under these conditions, about 30 min being required for a single run. Furthermore, the 5.8S rRNA from rat liver cells was eluted at a different position compared with that of the yeast, Saccharomyces cerevisiae, although they have the same nucleotide chain length but at different sequence. The 5.8S rRNA from the cellular slime mold, Dictyostelium discoideum, was eluted at the same position as that of yeast.