The compactization of a single DNA molecule in polyethylene glycol (PEG) solution was investigated both theoretically and experimentally. A theory is proposed taking into account the polyelectrolyte effect and redistribution of PEG within DNA coils. This approach makes it possible to describe the dependence of critical value, c, of PEG concentration at the point of DNA collapse on the degree of PEG polymerization, P, and on the concentration of low‐molecular salt, ns. Observation of single DNA molecule in solution of PEG has been carried out by means of fluorescence microscopy which allows one to observe the conformation of individual DNA directly. Direct evidence that the coil–globule transition of DNA occurs as first order phase transition was obtained. It was confirmed that the critical concentration of PEG decreases with an increase of the degree of PEG polymerization and salt concentration. The width of the coexistence region of coil and globule was found to be dependent on salt concentration and degree of polymerization of PEG. It was found that DNA undergoes re‐entrant globule–coil transition in concentrated solution of high‐molecular weight PEG. These experimental results correspond well to the theoretical predictions.

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