A sequential extraction method for RNA from rabbit liver.
When naphthalene-1,5-disulfonate (NDS) and phenol-cresol were used for extraction of rRNA from rabbit liver approximately 80% of the extractable RNA was recovered. By subsequent treatment of the resulting insoluble interface with 4-aminosalicylate and triisopropylnaphthalenesulfonate (PAS-TIPNS) the remaining RNA was extracted. Subcellular fractionation and polyacrylamide gel electrophoresis or sucrose density gradient analysis showed that this last fraction contrained 28s RNA from membrane-bound polysomes. Applying the sequential extraction method for isolation of poly(A)-containing mRNA showed that only a minor part of that RNA was extracted with NDS while the rest was recovered by PAS-TIPNS treatment of the interface. Thus, a purification of poly(A)-containing mRNA was possible when the sequential extraction method was used.
D000636:Aminosalicylic Acids / Q000494:pharmacology; D000818:Animals; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000032:analysis*; D008862:Microsomes, Liver / Q000032:analysis; D009282:Naphthalenesulfonates / Q000494:pharmacology; D011132:Polyribosomes / Q000032:analysis; D012313:RNA / Q000302:isolation & purification*; D012335:RNA, Ribosomal / Q000302:isolation & purification*; D011817:Rabbits; D013347:Subcellular Fractions / Q000032:analysis
D000636:Aminosalicylic Acids; D009282:Naphthalenesulfonates; D012335:RNA, Ribosomal; D012313:RNA