Understanding the abundance and distribution of massive jellyfish is necessary to forecast where or when their blooms will happen. The acoustic technique is one of the most useful methods of obtaining information if the acoustic characteristics of the targets are known. This study was conducted to determine the acoustic target strength (TS, dB) of two jellyfish species, Aurelia aurita and Cyanea nozakii, in the southern coast of Korea. For the ex situ measurements, 120, 200, and 420 kHz split beam transducers were used, and jellyfish with various bell lengths were arranged to prepare multiple jellyfish. Under 2 vertical individuals, the mean TS for multiple A. aurita at 120, 200, and 420 kHz was -72.7, -71.7, and -68.2 dB, respectively. In the case of 5 vertical individuals, the mean TS of the species was -71.3, -68.2, and -62.0 dB. Finally, the mean TS of C. nozakii at 120, 200, and 200 kHz was -62.0, -60.3, and -58.2 dB under 2 individuals and -58.1, -57.4, and -54.0 dB under 4 individuals, respectively. For both species, higher numbers of jellyfish resulted in a higher TS. In addition, higher frequencies were associated with a higher TS for the same jellyfish. These TS results for two species can be used as essential data for the acoustic detection of jellyfish in an open ocean or coastal area.