Substantial work has been conducted to estimate the river length required for recruitment of invasive Grass Carp Ctenopharyngodon idella and bigheaded carps (Bighead Carp Hypophthalmichthys nobilis and Silver Carp H. molitrix); however, the distance upstream and size of stream required for spawning remain unclear. Adult Grass Carp are regularly captured in Harry S. Truman Reservoir, Missouri, although there have been only unconfirmed sightings of Silver Carp. Fertile diploid Grass Carp are used legally for vegetation control in Missouri; however, it is uncertain whether Grass Carp in Harry S. Truman Reservoir are the result of natural reproduction or escapees. We used ichthyoplankton tows and environmental DNA (eDNA) to investigate which carp species were present in the reservoir basin and to determine whether these carps were spawning in reservoir tributaries. Grass Carp eggs were collected in four of five tributaries during June 2014. We did not detect eggs, larvae, or eDNA of Bighead Carp or Silver Carp. The occurrence of Grass Carp eggs generally coincided with rapid increases in discharge and turbidity. These tributaries may be the smallest rivers worldwide in which spawning of Grass Carp has been documented. We infer from the developmental stages captured and the distance from the reservoir that sufficient river length exists for egg survival. These results have broad implications concerning the establishment of Grass Carp in other novel habitats, including other reservoirs and lake basins, smaller rivers, and the Laurentian Great Lakes ecosystem.