Conditions of Phenol Extraction of RNA From Rat Liver Subcellular Cytoplasmic Fractions II. Effect of a Glycine NaOH Buffer on the Separation of Polysomal RNA

Abstract

A suspension of rat liver ribosomes in glycine NaOH buffer, pH 10. 5, with phenol was prepared after manually shaking for 30 sec with rapid strokes. The RNA was clarified immediately after by centrifugation. This extraction procedure was at 2°C and gave a high yield in RNA. Previous investigators omitted the addition of glycine NaOH buffer, thus preparing their suspension in approximately 10 min at 37°C⁴. Protein determinations and the integrity of RNA on gradients were used to determine the purity and stability of the RNA. A comparison was made between preparations of ribosomal RNA and microsomal RNA. The results showed that the former were much more stable.