Abstract
Methyl albumin kieselguhr (MAK) chromatography has been employed to separate ribosomal RNA from the rapidly labeled polyribosome-associated RNA (messenger RNA). Preliminary experiments indicated that the messenger RNA fraction which was used for MAK column analysis was free of any detectable contamination by nucleoplasmic heterogeneous RNA. Both messenger and nucleoplasmic RNA exhibited similar elution characteristics from a MAK column. The messenger RNA fractions recovered from the column were heterodisperse, sedimenting from 10 to 50S in a sucrose gradient. The nucleoplasmic RNA fractions were even larger. Evidence is presented which suggests that secondary structure, rather than base composition, of the RNA influences the separation of RNA on MAK columns. With the use of MAK columns it was possible to follow the kinetics of labeling of messenger and nucleoplasmic RNA in the presence of normal rates of ribosomal RNA synthesis. This column procedure offers an efficient method for preparing bulk quatities of eucaryotic messenger RNA for various metabolic and sequence studies.
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