A Liquid Chromatographic Preparation of Retroviral RNA

Liquid chromatography was used to prepare native dimer RNA from Moloney murine leukemia retrovirus by gel filtration on a TSK 6000PW column. Three RNA peaks were separated from viral lysate. RNA from the first eluted peak migrated in gel electrophoresis as a native dimer prepared by phenolic extraction and saccharose gradient separation. The last eluted RNA peak likely represents tRNA for proline. HPLC preparation was twice as fast and 20 times more productive than the other method, considering the quantity of pure RNA obtained for the same volume of viral lysate. Using several natural RNAs, it was verified that the dispersion coefficient was inversely proportional to RNA size, at least between 3.6 and 16.6 kb. Within the range of laboratory use, peak surface was in a direct ratio to the injected quantity of a given RNA species. Thus size exclusion chromatography could represent a valuable tool for preparation, analysis, and quantitation of large RNAs.