The Dayton Project was a research and development project to produce polonium during World War II, as part of the larger Manhattan Project to build the first atomic bombs. Work took place at several sites in and around Dayton, Ohio. Those working on the project were ultimately responsible for creating the polonium-based modulated neutron initiators that were used to begin the chain reactions in the atomic bombs.

The Dayton Project began in 1943 when Monsanto's Charles Allen Thomas was recruited by the Manhattan Project to coordinate the plutonium purification and production work being carried out at various sites. Scientists at the Los Alamos Laboratory calculated that a plutonium bomb would require a neutron initiator. The best-known neutron sources used radioactive polonium and beryllium, so Thomas undertook to produce polonium at Monsanto's laboratories in Dayton. While most Manhattan Project activity took place at remote locations, the Dayton Project was located in a populated, urban area. It ran from 1943 to 1949, when the Mound Laboratories were completed in nearby Miamisburg, Ohio, and the work moved there.

The Dayton Project developed techniques for extracting polonium from the lead dioxide ore in which it occurs naturally, and from bismuth targets that had been bombarded by neutrons in a nuclear reactor. Ultimately, polonium-based neutron initiators were used in both the gun-type Little Boy and the implosion-type Fat Man used in the atomic bombings of Hiroshima and Nagasaki respectively. The fact that polonium was used as an initiator was classified until the 1960s, but George Koval, a technician with the Manhattan Project's Special Engineer Detachment, penetrated the Dayton Project as a spy for the Soviet Union.

In December 1942, during World War II, Charles Allen Thomas, a chemist and director of research at Monsanto in St. Louis, joined the National Defense Research Committee (NDRC) as the deputy chief of its Division 8, which was responsible for propellants, explosives and the like. Early in 1943, he traveled to the east with Richard C. Tolman, a member of the NDRC, and James B. Conant, the president of Harvard University and the chairman of the NDRC, to witness a demonstration of a new underwater explosive. Conant and Tolman took the opportunity to quietly investigate Thomas' background. He was then invited to a meeting in Washington D.C., with Brigadier General Leslie R. Groves, Jr., the director of the wartime Manhattan Project responsible for building an atomic bomb. When he got there, Thomas found Conant was also present.

Groves and Conant were hoping to harness Thomas's industrial expertise. They offered him a post as a deputy to Robert Oppenheimer, the director of the Los Alamos Laboratory in New Mexico, but he did not wish to move his family or give up his responsibilities at Monsanto. Instead, he accepted the role of coordinating the plutonium purification and production work being carried out at Los Alamos, the Metallurgical Laboratory in Chicago, Radiation Laboratory in Berkeley, and Ames Laboratory in Iowa. Chemistry and metallurgy at Los Alamos would be led by the youthful Joseph W. Kennedy.

At Los Alamos, physicist Robert Serber proposed that instead of relying on spontaneous fission, the chain reaction inside the atomic bomb should be triggered by a neutron initiator. The best-known neutron sources were radium-beryllium and polonium-beryllium. The latter was chosen, as polonium has a 138-day half-life, which made it intense enough to be useful but not long-lived enough to be stockpiled. Thomas took charge of the development of techniques to industrially refine polonium for use with beryllium in the "urchin" internal neutron initiators. This effort became the Dayton Project.

Thomas brought in key personnel from Monsanto's Thomas and Hochwalt Laboratories in Dayton, Ohio, including Caroll Hochwalt, James Lum and Nicholas Samaras. Thomas became Director of the Dayton Project, with Hochwalt as Assistant Project Director and Lum as Laboratory Director. They decided that about twelve chemists would be required, and Lum set about recruiting professors, graduate students and industrial chemists from universities and laboratories in the area. The first of these recruits commenced in August 1943, but few had any experience with radiochemistry. Numbers increased from 46 full-time employees at the end of 1943 to 101 at the end of 1944, 201 at the end of 1945, and 334 at the end of 1946, including 34 members of the Army's Special Engineer Detachment.

Office space was initially found in the Monsanto offices at 1515 Nicholas Rd, which became known as Unit I. Unit II was the Monsanto Rocket Propellant works off Betty Lane near Ohio State Route 741. While it was administered by Monsanto, it was not used by the Dayton Project. The site handled explosives including ammonium nitrate and ammonium picrate, but no radioactive materials were handled there. Work at Unit II ceased in the fall of 1945. Consideration was given to using it in December 1946, but this proposal was rejected in favor of erecting a Quonset hut at Unit III.