Dietary studies of polyps and ephyrae are important to understand the formation and magnitude of jellyfish blooms and provide important insights into the marine food web. However, the diet of polyps and ephyrae in situ is largely unknown. Here, prey species of the polyps and ephyrae of the moon jellyfish Aurelia coerulea in situ were identified using high-throughput DNA sequencing techniques. The results show that A. coerulea polyps and ephyrae consume a variety of prey items. The polyps consume both planktonic and benthic prey, including hydromedusae, copepods, ciliates, polychaetes, stauromedusae, and phytoplankton. A. coerulea ephyrae mainly feed on copepods and hydromedusae. Gelatinous zooplankton, including Rathkea octopunctata and Sarsia tubulosa, were frequently found as part of the diet of A. coerulea polyps and ephyrae. The utilization of highthroughput sequencing technique is a useful tool for studying the diet of polyps and ephyrae in the field, complementing the traditional techniques towards a better understanding of the complex role of gelatinous animals in marine ecosystems.