Direct purification of polyadenylated RNAs from isolated polysome fractions.
We report a simplified and improved method for obtaining polyadenylated RNAs (poly(A) RNAs) from polysome fractions. Isolated polysomes were subjected directly to poly(U)-Sephadex column chromatography without conventional purification of polysomal RNAs by phenol extraction followed by ethanol precipitation. The yield of poly(A) RNAs by this direct purification method was about twice that obtained by the conventional method. When the two poly(A) RNA preparations were used in two-dimensional polyacrylamide gel electrophoretic analysis of cell-free translation products and cDNA synthesis, biological activity of the directly purified poly(A) RNA was equal to or even better than that of conventionally purified poly(A) RNA.
D000818:Animals; D002474:Cell-Free System; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D011061:Poly A / Q000302:isolation & purification*; D011132:Polyribosomes / Q000032:analysis*; D014176:Protein Biosynthesis; D012313:RNA / Q000302:isolation & purification*; D012333:RNA, Messenger; D011817:Rabbits
D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D004247:DNA