The invention discloses a palmtop pyrosequencing system based on digital microfluidic technology, and belongs to the technical field of digital microfluidics. A digital microfluidic chip, an electromagnetic module, and an integrated circuit are connected, and the integrated circuit and a chemiluminescent collection module are connected to a computer. According to the palmtop pyrosequencing systembased on digital microfluidic technology, a printed circuit board is used as a chip substrate, and based on an on-chip dielectric wetting principle, the electrifying sequence of electrodes that are located on the chip is regulated to achieve liquid drop generation, transportation, mixing and splitting in a sequencing process. The system is easy to operate and is miniaturized, and the produced optical signal can be fast rapidly read, so that single base on the chip is measured. The system is simple in structure, low in cost, easy to integrate, and high in detection sensitivity, the sequencing cost is greatly reduced, and the system can be widely applied to the field of pyrosequencing-based gene detection.