Ben fights Dr. Animo, who has kidnapped Gwen and Grandpa Max, and threatens to release his DNA bomb and de-evolutionize the world. Ben (as Heatblast, XLR8, or Eye Guy in different beginnings) rescues Grandpa and Gwen, defeats Animo's latest invention, and sets off an error in the bomb by overloading the system. A DNA wave blows through and damages the Omnitrix. Ben changes into Grey Matter and knocks out Dr. Animo. Gwen, Ben and Grandpa go to the shopping mall but the Omnitrix releases a sudden energy burst, banning them from the mall.
Tetrax senses the Omnitrix and comes to Earth to warn Ben about it. The watch is in self-destruct mode and will destroy itself along with Ben (and the universe, but Tetrax does not reveal this until later). Ben, Tetrax, and his pilot, Gluto, travel across the galaxy to find Azmuth, the creator of the Omnitrix. Gwen soon appears on the ship to help Ben. They find the DNA signature of Azmuth in the Omnitrix and find that he is on Incarcecon, the prison planet.
After some time of searching on Incarcecon, Ben (in a costume with Gwen) thinks he spots Vilgax. He changes into Perk Upchuck and fights him, only to reveal that "Vilgax" is a female Chimera Sui Generis named Myaxx, who claims she created the Omnitrix. Ben, Gwen, and Tetrax get themselves into a street-fight. After escaping the fight, she reveals she was Azmuth's assistant and directs Ben and the others to Xenon, where Azmuth lives. Their ship was intercepted and boarded by Vilgax and his drones, but they were stopped by Ben, Gwen, Tetrax, Gluto, and Myaxx. In that action, Gluto was assumed dead.
When they got to Xenon, they were attacked by a group of wild Floraunas, where Gwen was captured by them and assumed dead. After Ben's sadness, Tetrax told him the tragic story about him bringing Vilgax the final element he needed to destroy his home planet, Petropia. Even though transforming would accelerate the self-destruct, Ben transformed into Cannonbolt to break into Azmuth's lab, but Azmuth refused to help them. Ben then sees Vilgax, Sixsix, and their massive army of drones. Azmuth removed the main part of the Omnitrix, but Ben, Tetrax, and Myaxx still fought.
Soon, it was revealed that Gwen and Gluto were still alive and helped battle. Azmuth then fixes the Omnitrix and unlocks Way Big. Ben then transforms into Way Big, defeats Vilgax's army of drones, grabs Vilgax, and throws him into space.
Ben is brought back to Earth, finds out that zombies are attacking the mall, and heads off to stop them.
Tetrax: Don’t worry, I’ve had many of these tests performed on myself. Never felt a thing.
Ben: But don’t you have like diamond hard skin?
Tetrax: ...Good point. Let's get started!
Ben: (Transforms into Way Big for the first time) Whoa! Check me out! I'm not just big! I'm WAY Big!
Vilgax: Release me or suffer the consequenses!
Way Big: Pretty big talk for such a little guy. Vilgax: (Scratches Way Big's hand) Way Big: OW!! THAT'S IT!!! YOU ARE SO OUTTA HERE!!!! (Throws him into space)
Vilgax: NOOOOOOOOOO!!!!!!!
Azmuth: You coming?
Myaxx: Are you going to give me some respect from now on? Azmuth: Meh. I wouldn't count on it.
Myaxx: Guess it is better than that prison planet.
When Ben and Gwen are fighting aliens on Incarcecon, right before the Tetramand destroys Tetrax's hoverboard, the Omnitrix is missing from Ben's wrist.
When Azmuth is re-inserting the Omnitrix dial, the "tower" of the dial is grey and green, as opposed to the standard black and green.
Tetrax calls Wildmutt a "Vulpin" instead of a "Vulpimancer".
In the Polish version of this movie, Gwen said "Heatblast!" to Ben, who was XLR8, and in the Arabic version, XLR8 said to Animo's insect that he would bake him like a potato, despite the fact that he was not Heatblast.
The controls that Tetrax uses to scan the Onmitrix for the DNA signature is almost identical to the computer that helps the "Precrime" officers in 'Minority Report'.
This movie was intended to be the Series Finale of the original Ben 10 Series.
Ben 10: Secret of the Omnitrix was three episodes put together into one movie.