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After looking at the thread title of the one in the Bioshock section, I figured we might as well put one up for any game.
Mine would have to be in Starcraft, when Kerrigan starts to get over-run by the Zerg, and she starts talking about how she needed immediate evac, and Mengsk just takes off, and Raynor gets very pissed off. Always going to be the pinnacle of story/acting in a video game, to me. I'll never forget this scene.