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We observed single DNA molecules at different ethanol concentrations by using fluorescence microscopy. Large single DNA molecules undergo reentrant conformational transitions from elongated coil into folded globule and then into elongated coil state, accompanied by the increase of the concentration of ethanol in a low‐salt aqueous environment. The second transition from globule into the coil state occurs at around 70 % (v/v) ethanol. From circular dichroism (CD) measurements, it is confirmed that the reentrant transition of the higher order structure proceeds together with the transitions of the secondary structure from B to C and, then, from C to A in a cooperative manner. The determined mechanism of the reentrant transition is discussed in relation to the unique characteristics of solutions with higher ethanol content, for which clathrate‐like nanostructures of alcohol molecules are generated in the surrounding water.

Highly Concentrated Ethanol Solutions: Good Solvents for DNA as Revealed by Single‐Molecule Observation