Holzhauer won $80,615 on Thursday's episode, bringing his total to $1,608,627.
The final clue from the night was: "A character in this 1944 play is said to be like a piece of her own collection, 'too exquisitely fragile to move from the shelf.'"
The correct answer was "What is 'The Glass Menagerie?'"
The 34-year-old from Las Vegas is already No. 2 on the all-time regular season cash winnings list. He also holds the "Jeopardy!" record for single-day cash winnings.
Outside of the other contestant's he'll face, Holzhauer's long-term challenge is beating Ken Jennings: the only person who remains in the top spot for both consecutive wins and cash earnings.
And it could take a while. Jennings won 74 games and cashed out at $2,520,700.
Holzhauer uses a controversial method known as the "Forrest Bounce," which has helped him get so far on the show. With this strategy, a contestant jumps from category to category, choosing tiles out of order to throw off opponents.
Holzhauer's next game is Friday.
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