Separation of newly-synthesized RNA by organomercurial agarose affinity chromatography.
Mouse myeloma cells, MOPC-31C, were incubated in the presence of 2-thiouridine and newly-synthesized RNA which appeared to contain 2-thioUMP as a constituent was separated from preexisting RNA by affinity chromatography using organomercurial agarose as a support. Both pH and salt concentration greatly affected the specific adsorption of the newly-synthesized RNA on the column. Under optimal conditions the rate of adsorption of the newly-synthesized RNA on the column was proportional to the logarithmic concentration of 2-thiouridine in the culture medium. Furthermore, at a given concentration of 2-thiouridine in the medium, a shorter incubation period caused a reduction of the rate of RNA adsorption on the column. The molecular size distributions of both total RNA and the adsorbed fraction, synthesized during 30 min in the presence of 2-thiouridine, were similar to that of RNA synthesized in the absence of the drug.
D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009941:Organomercury Compounds; D009994:Osmolar Concentration; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000302:isolation & purification; D012965:Sodium Chloride; D013891:Thiouridine / Q000378:metabolism
D009941:Organomercury Compounds; D012334:RNA, Neoplasm; D013891:Thiouridine; D012965:Sodium Chloride