Ainslie's dream undefeated run this season came to a deflating end in the AFL Canberra first-grade women's grand final on Saturday evening.
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Queanbeyan celebrated an impressive double as the women backed up the men's premiership triumph in the earlier game at Phillip Oval, to defeat the favourites Ainslie 6.4 (40) to 5.5 (35).
"I don't think the double has been done in a while, certainly not in the seven years I've been here," Tigers coach Chris Clifton said in his final game with the club before he relocates to Victoria.
"The women's and men's teams are so close and this will only bring the club even closer.
"We knew we could push Ainslie, we just needed to turn up with the right attitude and for a few bounces to go our way - which they did.
"It's the last game I'm coaching so it's a fairytale finish."
The Tricolours took the early lead through Rachel Friend, the competition's leading goalscorer, but it was the Tigers that slowly took control of the match.
Sophia Arcus levelled the score before a Ruby Grounds first-quarter double - that included a beauty of a right-foot banana - saw the Tigers take the lead.
Friend kept Ainslie in touch with her second of the night, as the Tricolours trailed by a goal going into the second term.
Queanbeyan threatened to run away with the game extending their lead to eight points at the half, 4.3 (27) to 3.1 (19).
However Ainslie had picked up some momentum toward the end of the quarter and could have reduced the deficit further if Maddison Holloway had collected a mark 30 metres out a millisecond earlier, just beaten by the siren to deny her a shot at goal.
After the break, the Tricolours had the wind in their sails and continued to flex their muscles in the stoppages and in defence, keeping the Tigers scoreless in the third quarter.
At the other end of the field they kept wave after wave of attack coming before a momentous goal for veteran Tricolours star Dani Curcio.
In the game that sealed Ainslie's ticket to the grand final a fortnight ago, the Tricolours legend broke the all-time games played at the club and for that achievement the Alan Ray Oval scoreboard was named after her.
In that game she was also concussed, but was able to return for the season decider, and in her very first grand final appearance since playing with the club since 2001, Curcio scored a memorable goal.
Going into the final quarter, Ainslie held a one point lead, but it was the Tigers who got their noses in front with a few pivotal plays.
In a stunning moment that would make Shane Warne proud, Arcus' kick inside 50 was untouched when it bounced nearly 90 degrees, over the heads of Ainslie defenders and through the posts for an important score.
The Tricolours had chances late but failed to convert, while the Tigers were making their opportunities count.
Best on ground Ella Ross' determined sprint to pick up the crumbs and snap from close range extended their lead to 10 points before India Lehman added a behind.
Renee Friend put Ainslie within five points with 20 seconds left but it was all too late as the final siren sounded, and Queanbeyan claims victory.
"Queanbeyan put on an awesome show today," Ainslie's Holloway said after the loss.
"Our girls had an incredible season and I'm so proud of every single one of them."
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