Research on nanofiber materials is actively done around the world today. Various types of nanofibers have been synthesized using an electrospinning technique. The most important component when synthesizing nanofibers using the electrospinning technique is a DC high voltage power supply. Some requirements must be fulfilled by the high voltage power supply, i.e., it must be adjustable and its output voltage reaches tens of kilovolts. This paper discusses the design and development of a high voltage power supply using a diode-split transformer (DST)-type high voltage flyback transformer (HVFBT). The DST HVFBT was chosen because of its simplicity, compactness, inexpensiveness, and easiness of finding it. A pulse-width modulation (PWM) circuit with controlling frequency and duty cycle was fed to the DST HVFBT. The high voltage power supply was characterized by the frequency and duty cycle dependences of its output voltage. Experimental results showed that the frequency and duty cycle affect the output voltage. The output voltage could be set from 1 to 18 kV by changing the duty cycle. Therefore, the nanofibers could be synthesized by employing the developed high voltage power supply.