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Put it at the end of a reveal sequence, after the twist is shown and before the next scene begins. Pair it with a dramatic dun dun dun sting beforehand to build suspense, then follow it with a vine boom for impact once the villain takes over.
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This is a roughly six-and-a-half-second villain laugh tied to Omega Flowey, the distorted final boss form in Undertale. The cackle is high, mocking and deliberately overblown, stretching from smug amusement into something openly threatening. It makes a scene feel tense, creepy and out of control, like a plan has just turned against everyone in it. The laugh comes from the 2015 indie role-playing game created by Toby Fox.
