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Synchrotron radiation circular dichroism revealed that the overall CD profiles of a 21\u2010mer and a d(CG)<jats:sub>9<\/jats:sub> duplex remain similar in the simulated crowded fluids compared with those in Tris buffer, but the amplitude ratios of individual maxima and minima vary with the concentration of DNA and the media, suggesting possible conformational changes. Using fluorescence resonance energy transfer, this work confirmed that hairpin and duplex formations are not affected by the crowding condition. 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