DOTA may refer to:
- Defense of the Ancients, a custom scenario for the video game Warcraft III.
- Dota 2, the stand-alone sequel to the previously listed game, developed on the Source engine.
- Vi sitter i Ventrilo och spelar DotA, a song by Basshunter based on the previously listed game.
- DOTA (chelator), a chemical.
- DOTA-TATE, a substance used in cancer treatment.
- 90Y-DOTA-biotin, another substanec used in cancer treatment.
- Dota, a canton in the province of San José, Costa Rica.