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Background interactions,including macromolecular crowding,confinement,and adsorption,[1] are able to greatly influence the equilibria and rates of reactions in which they participate.DNA nanotube assembled by DNA tiles can be classified as a novel type of living polymer.[2] In this work,it serves as a model system to study the effect of macromolecular crowding and the confinement on the assembly process.Polyacrylamide (PAM),which has no interactions with DNA,is employed to prepare the solutions of varying concentrations.Single molecular fluorescence microscope is used to monitor the real-time assembly of DNA nanotube in PAM solution.