This work proposed a novel polymerase chain reaction (PCR) enhancer and testified its function to restore and enhance the fluorescence signal of dsDNA binding dyes for PCR performed on digital microfluidic (DMF) platforms. The dsDNA binding dyes such as SYBR Green I (SGI) have been widely used as amplification indicators in quantitative real-time PCR. However, the reduced sensitivity and the false negative signal of on-chip PCR make them incompatible with microfluidic systems. With its unique structure, the proposed PCR enhancer solved this problem and enhanced the signal of on-chip PCR to a large extent.