Place it at the end of a failed-plan sequence, after the setup and just before the reaction shot. Pair it with It Was At This Moment He Knew for the build-up, then follow it with Fail Sound to close the joke, or swap in Darth Vader Nooo for a second villain-style reaction.
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This 3.5-second clip is Doctor Octopus letting out a long, desperate 'Noooooo' as his plan collapses in Spider-Man 2 (2004). The stretched cry sounds theatrical and over-the-top, turning a villain's defeat into something funny and strangely satisfying to hear again.