Condensin plays a central role in the organisation of chromosomes by compacting chromatin into loops during mitosis. Condensin achieves this through a loop extrusion mechanism that remains poorly understood. To identify the molecular steps of yeast condensin during loop formation, we used optical tweezers with fluorescence detection. We find that single yeast condensin complexes use ATP to extrude DNA through distinct 15 nm steps, thus advancing ~45 base pairs (bp) per step. Under increasing load, the condensin step size remains constant while step-dwell times increase, and stalls at forces >1 pN. We also show that nucleosome arrays hinder processive condensin extrusion and demonstrate that the histone chaperone FACT is required for compaction of nucleosomal arrays by condensin. Importantly, FACT-assisted compaction on nucleosomes also occurs through distinct 15 nm steps. Finally, we show that FACT is required for correct condensin localisation in vivo. Our results establish that loop extrusion by yeast condensin involves a 45 bp stroke that requires FACT for condensin function on chromatin

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