Uchiyama S, Imamura T, Nagai S, Konishi K
J Biochem. 1981 Sep;90(3):643-8. doi: 10.1093/oxfordjournals.jbchem.a133518.
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.
通过使用基于硅胶的水性凝胶柱TSK-GEL G 300 SW的高速凝胶过滤方法,在含有0.1%十二烷基硫酸钠(SDS)的pH 7.0的200 mM磷酸钠缓冲液中,三种低分子量RNA种类,即5.8S和5S核糖体RNA(rRNA)以及转运RNA(tRNA),彼此之间以及与高分子量RNA种类清晰地分离。在这些条件下获得了出色的重现性和分辨率,单次运行大约需要30分钟。此外,尽管大鼠肝细胞的5.8S rRNA与酿酒酵母具有相同的核苷酸链长度但序列不同,但其洗脱位置与酵母的不同。来自细胞黏菌盘基网柄菌的5.8S rRNA与酵母的在相同位置洗脱。