A single radio frequency bucket of the 88-Inch Cyclotron is filled using a fast chopper located in the axial line. The bucket then accelerates until it reaches the deflector, at which point, it is extracted as a train of bunches. This phenomenon can be attributed to the characteristic multi-turn extraction of the cyclotron and, by simplifying the complex dynamics of a cyclotron, corresponds to the conceptual transfer function of the cyclotron. The confirmation of the single radio frequency bucket injection was achieved by operating the cyclotron in the third harmonic mode and observing the absence of intermediate bunches during the multiple-bunch extraction.