An efficient, rapid procedure for the isolation of polysomes from Novikoff hepatoma ascites cells that avoids mechanical shear was developed, and Ivory was used as the detergent to lyse the cells. This method yields classes of large polysomes and 3- to 5-fold increased yields over that of routine homogenization procedures. Up to 70% of the polysome population is larger than tetramers. The polysomes and ribosomes are morphologically intact and contain undegraded ribosomal RNA and biologically active messenger RNA. This procedure also yields nuclei free of cytoplasmic contamination as a by-product of polysome isolation. These nuclei do not show evidence of protein degradation and are enzymatically active with respect to RNA polymerases I and II.
These studies were supported by Cancer Research Center Grant CA-10893, P.5 and Training Grants CA-5154 and GM/670 from the NIH.