High-speed preparative-scale separation and purification of ribosomal 5S and 5.8S RNA's via Sephacryl S-300 gel filtration chromatography.
In this work we present a rapid and economical alternative to Sephadex and high performance size exclusion chromatography (HPSEC) for preparative-scale separation and purification of low molecular weight RNA's: 5.8S RNA, 5S RNA, and tRNA's. These three RNA species can be well resolved from each other and from higher molecular weight RNA species via Sephacryl S-300 gel filtration chromatography under mild eluting conditions: 10 mM Tris-HCl buffer, pH 7.5, containing 1.0 M NaCl. For a sample load of about 250 mg, the resolving power of a Sephacryl S-300 column (78 X 3.2 cm) is comparable to that of a 4.5 times larger Sephadex G-75 column (144 X 5 cm). Moreover, the total separation period is 2.5 times shorter for the Sephacryl method. Up to 500 mg or more of crude ribosomal RNA mixtures could be separated via two Sephacryl S-300 columns operated in tandem.
D000180:Acrylic Resins; D002850:Chromatography, Gel / Q000379:methods; D008970:Molecular Weight; D012331:RNA, Fungal / Q000302:isolation & purification; D012335:RNA, Ribosomal / Q000302:isolation & purification*; D012440:Saccharomyces / Q000235:genetics
D000180:Acrylic Resins; D012331:RNA, Fungal; D012335:RNA, Ribosomal; C037239:Sephacryl Superfine