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We confirm Landauer’s 1961 hypothesis that reducing the number of possible macroscopic states in a system by a factor of two requires work of at least kT ln 2. Our experiment uses a colloidal particle in a time-dependent, virtual potential created by a feedback trap to implement Landauer’s erasure operation. In a control experiment, similar manipulations that do not reduce the number of system states can be done reversibly. Erasing information thus requires work. In individual cycles, the work to erase can be below the Landauer limit, consistent with the Jarzynski equality.

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