Development of radio frequency identification (RFID) technology and passive integrated transponder (PIT) tags has substantially increased the ability of researchers and managers to monitor populations of aquatic organisms. However, use of transportable RFID antenna systems (i.e., backpack-mounted) is currently limited to wadeable aquatic environments (<1.4 m water depth). We describe the design, construction, and evaluation of a boat-mounted RFID antenna to detect individually PIT-tagged benthic aquatic organisms (mussels). We evaluated the effects of tag orientation on detection distances in water with a 32-mm half-duplex PIT tag. Detection distances up to 50 cm from the antenna coils were obtained, but detection distance was dependent on tag orientation. We also evaluated detection distance of PIT tags beneath the sediment to simulate detection of burrowing mussels with 23- and 32-mm tags. In sand substrate, the maximum detection distance varied from 3.5 cm and 4.5 cm (vertical tag orientation) to 24.7 cm and 39.4 cm (45° tag orientation) for the 23- and 32-mm PIT tags, respectively. Our results suggest a 1.4-m total detection width for tagged mussels on the substrate surface by the boat-mounted antenna system regardless of tag orientation. However, burrowed mussels may require multiple passes to increase detection that would be influenced by depth, tag orientation, and tag size. Construction of the boat-mounted antenna was relatively low in cost (<500 USD) and had several advantages (less labor and time intensive, increased safety) over traditional mussel sampling techniques (diving, snorkeling) in nonwadeable habitats.