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The demonstration' that Q8-replicase2 can effect the synthesis of biologically competent copies, and that the RNA is the instructive agent,3 provided a system which for the first time permitted an unambiguous in vitro analysis of a replicating mechanism. Examination of the early course of the reaction revealed4 the existence of a sixminute latent period which precedes the appearance of the first new infectious strands and which is accompanied by an "eclipse" of the initiating templates as infectious entities. A detailed analysis5-7 of the early events in the latent period was made possible by the use of electrophoretic separation in acrylamide gels.8' 9 In these experiments the input RNA was labeled with H3 and the newly synthesized product with p32, permitting a simultaneous examination of both. The data obtained indicate that all of the first product synthesized is initially associated with H3-templates in complexes possessing the properties of Hofschneider'0 structures (HS). Subsequently Franklin" structures (FS) appear containing template and product. Soon after the formation of FS complexes, mature Q13-RNA molecules are found. Further insight into the nature of the Qf3-replicase reaction has been gained from the demonstration'2 that guanosine 5'-triphosphate (GTP) is the first subunit added. This may be related to the poly G polymerase observed'3 14in Q,8-replicase preparations, an activity which has been shown'4 to be associated with one of the two protein components required for complete synthetic function by Qf3-replicase. The rapidity with which information is being generated by study of the Qf3replicase reaction makes it desirable to begin a more detailed chemical analysis of the Qi3-RNA molecule, with the ultimate aim of obtaining its complete sequence. Partial degradation of RNA with endonucleases has been used in studies leading to the elucidation of the primary structure of sRNA.'5' 16 This technique has also been applied to the fragmentation of ribosomal RNA's;"'-19 and of 5S RNA using20 alkylated ribonuclease. In principle these procedures can be applied to any homogeneous population of RNA molecules. We report here a technique for the production of two specific fragments from the RNA of the coliphage QB.21 The fragments have been characterized and their orientation with respect to the originalRNA strand determined.

Specific cleavage of Qbeta-RNA and identification of the fragment carrying the 3'-OH terminus.