The legend of El Dorado originated with the Muisca people of Colombia, who migrated into the area around 500 BCE and formed a confederation that was, along with the Aztecs, Inca, and Maya, one of the four major civilizations of the ancient Americas.

The ancient Muisca idolized the Sun and Moon, as well as natural places such as lakes and caves. They thought gold represented creativity and cosmic energy, and sacrificed gold objects to the divine cosmic energies by throwing offerings, called tunjos, into Lake Guatavita. Tunjo offerings included sculptures of humans and animals made out of gold, often alloyed with silver and copper and sometimes studded with precious stones.

Muisca rulers, referred to as the zipa, made offerings to the water goddess Chie in an elaborate cleansing ritual, then covered themselves in gold dust and waded into the lake. A glimpse of this ritual by a European explorer is most likely where the origin of the legend of El Dorado began.