A digital microfluidic system based on electrowetting-on-dielectric (EWOD) technology has attracted considerable attention because of its promising potential in biological, biochemical and biomedical applications. The digital microfluidic (EWOD) system enables a discrete droplet to be manipulated through reconfigurable paths or channels defined by electrode actuation. Minute sample and reagent volume, precise droplet control, low energy consumption, low cost, rapid analysis, portablility for point-of-care diagnosis, flexibility for integrating with detector, multiple-step and simultaneous reaction processes are all advantages of EWOD system. Thus, it has become a promising and popular tool for bio-applications such as cell assays, proteomics, enzyme assays, immunoassays, DNA extraction, DNA repair and DNA amplification. In DNA-based applications, ordinarily, the DNA replication (e. g., DNA amplification) is necessary. Among the DNA replication techniques, Polymerase Chain Reaction (PCR) because of its simplicity and cost-effectiveness is the most common laboratory technique. Therefore, the development of EWOD-based microfluidic PCR system combined with other novel techniques for DNA amplification or DNA-based assays has extensively been reported. In this paper, the recent developments and studies of EWOD microfluidic system for DNA-based applications published in the last five years have been reviewed. (C) Koninklijke Brill NV, Leiden, 2011