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The two Americans have won 14 doubles titles at the majors and their hopes of extending it to 15 was dashed off on Wednesday by the Czech duo.
The loss leaves 40-year-old Serena free to focus on what is widely expected to be the final singles campaign of her landmark 27-year career before retirement.
On Friday, the 23-time Slam singles winner will face Australia's Ajla Tomljanovic for a place in the fourth round.
Serena and her 42-year-old sister went down fighting, clawing back from 1-4 down in the second set before the Czech pair steadied the ship.
"I'm still in shock," said Hradecka.
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