Autonomous motor: We constructed a bipedal autonomous DNA-motor with a coordinate activity between the two motor legs and monitored its activity using SMF techniques. The measurements are done in-situ which enables monitoring the motor’s progress and structural dynamics without disturbing its activity. Our kinetic measurements of the motor’s assembly and activity indicate that it takes dozens of seconds to complete reactions, rather than hours, if components are properly designed. However, we monitor side reactions that significantly reduce the yield of the reaction and resulting in defected motors. We are now working on implementing new strategies for motors’ preparation which will prevent side reactions, altogether, resulting in much higher yield. On the methodological side, we have measured the motor’s dynamics and its interaction with its energy source, a DNA-fuel, in equilibrium and non-equilibrium conditions. Our work demonstrates that by using SMF, one can construct a DNA-machine and monitor its activity in ways not possible with conventional methods. We demonstrate that our methods enable simultaneous in-situ monitoring of the motors efficiency, integrity and activity