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.