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We perform a single molecule force spectroscopy study to characterize the role of the size (molecular weight) and concentration of depleting agents on DNA condensation and on the stabilization of the double-helix structure, showing that important features such as the threshold concentration for DNA condensation, the force in which the melting plateau occurs and its average length strongly depend on the depletant size chosen. Such results are potentially important to understand how the presence of surrounding macromolecules influences DNA stabilization inside living cells and therefore advance in the understanding of the crowded cell environment on DNA-related functions.

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