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We present the results of high temperature 500 K Molecular Dynamicssimulations of the DNA i-motif. The essential dynamics and the main unfoldingpathways are compared to a biased metadynamics simulation at 300 K. Our resultsindicate a remarkable agreement of the concerted motion at both temperatures.The transition can be described by a few number of eigenvectors indicating asimple unfolding process. Two main mechanisms for the unfolding pathway at 500K can be detected which are in good agreement to the results of the biasedsimulation at 300 K.

High temperature unfolding simulations of a single stranded DNA i-motif