Cytoplasmic RNA from hen reticulocytes, mouse sarcoma 180 ascites cells, rat liver and barley embryos. Their preparation and purification by a standard procedure and characterization by polyacrylamide gel electrophoresis

Abstract

1. 1. This paper describes a standard procedure for the preparation and purification of RNA from the post-mitochondrial supernatants of a number of eukaryotes. 2. 2. Cytoplasmic RNA was fractionated by NaCl precipitation. The 28 S (26 S), 18 S and 5·8 S rRNA, and 9 S RNA, in the NaCl insoluble fraction were separated by a two-step sucrose gradient fractionation procedure. Poly(A)-containing mRNA in hen 9 S RNA was purified by affinity chromatography. The 5 S rRNA and tRNA in the NaCl-soluble fraction were fractionated by gel filtration. 3. 3. Polyacrylamide gel electrophoresis showed that the above RNA species were remarkably stable and homogeneous. Differences were found in the 26–28 S rRNA, 5·8 S rRNA, and 9 S RNA of different eukaryotes, but other cytoplasmic RNA species were identical.