A general method for determination of nucleotide sequences in nucleic acids

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Abstract
A method is proposed for the determiantion of nucleotide sequences in nucleic acids. Briefly, the procedure involves the formation of derivatives specific for either the 3′ or 5′ end of nucleic acid. This is followed by a partial enzymatic hydrolysis to yield specific fragments. The functions of the chemically modifying derivative are (a) to serve as a reference point which positions each base according to its distance from the modified end, (b) to provide a convenient means of separating oligomers with derivatives from those without derivatives, (c) to permit the use of additional parameters of fractionation of oligomers with derivative ends. The features of this method include: construction of long sequences from shorter fragments, unequivocal positioning of fragments, independence from quantitative procedures, economical use of nucleic acid polymer, determination of sequences from both ends simultaneously, application to DNA and RNA. Data are presented on the action of pancreatic ribonuclease on bis‐2,4 dinitrophenyl hydrazone derivatives of yeast ribonucleic acid and polyuridylic acid.