A new and versatile method for the incorporation of nucleotides, polynucleotides, and nucleic acids onto cellulose has been developed. The method involves specific activation of the terminal monosubsti-tuted phosphate or polyphosphate group of the nucleotide or polynucleotide (in aqueous solution at pH 6) in the presence of cellulose Chromatographic paper. The activation is brought about by N-cyclohexyl-N′-β-(4-methylmorpholinium)ethyl carbodiimide p-toluenesulfonate and the condensation with the cellulose is achieved by a novel technique in which the reaction mixture is concentrated within the fibers of the cellulose. The products of the reaction are considered to consist of nucleotides or polynucleotides connected to the cellulose by ester linkages between the terminal phosphate or polyphosphate groups of the nucleotide chains and the sterically favored hydroxyl groups of the cellulose. The reaction has been applied to a variety of nucleotides, polynucleotides, and nucleic acids, containing terminal phosphate, diphosphate, or triphosphate groups, and the yields of incorporation have varied from 15 to 71%. Polynucleotide celluloses prepared by this method are expected to be of some value in the study of those enzymes which are concerned with the synthesis or degradation of nucleic acids. In addition, the capacity of the method to readily incorporate nucleotides and polynucleotides onto an insoluble support should permit the development of new methods for the chemical and enzymatic synthesis of polynucleotides as well as new techniques for the fractionation and sequence analysis of polynucleotides from natural sources. © 1968, American Chemical Society. All rights reserved.
