Isolation and analysis of RNA formed by cell-free extracts from tobacco leaves

The RNA-like product obtained by incubation of ATP, GTP, UTP, and CTP with a cell-free extract from tobacco leaves has been purified by phenol extraction and passage through Sephadex. When either [α-³²P]ATP or [α-³²P]GTP was included in the reaction mixture the major portion of the incorporated counts remained associated with the RNA during purification. In both cases, alkaline hydrolysis of this purified, labeled product yielded AMP, GMP, CMP, and UMP all of which were ³²P-labeled. This provides direct evidence that the labeled product of the reaction contains all four nucleotides linked by interribonucleotide bonds to form a true RNA.