Place it at the peak of a fail, right after a shocked sound sets up the reaction and just before the Oh No No No Laugh releases the tension. That order builds the joke in three beats, with this clip as the centrepiece in the middle for maximum impact.
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This is a loud, drawn-out human voice stretching the word no into a long wail that lasts just over a second. The single shouted syllable rises and holds, sounding desperate and over the top. It feels funny and dramatic at once, like a cartoon refusal to accept bad news, and it works because the exaggeration is so immediate.