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The game includes over 170 puzzles.

Professor Layton and the Last Specter also includes an additional role-playing game entitled Professor Layton's London Life, available from the start of the game. London Life, in which players interact with various characters from the series in a town called "Little London", was advertised to contain over 100 hours of content. London Life was removed from the European versions of the game in order to prevent a significant delay of the game's release for translation.