A double from Brackley defender Pablo Mills sent Gainsborough crashing to a 3-2 defeat.
The former Rotherham and Derby defender punished a goalkeeping blunder from Jan Budtz with ten minutes remaining, after Trinity had come from two goals down to equalise.
Budtz came to claim a cross from a free kick, misjudged it and Mills was able to roll the ball into an empty net to move the Northamptonshire side out of the relegation zone and to within two points of Gainsborough.
The result also added to the ‘bogey team’ status of Brackley as the Saints have now defeated the Blues in their last four meetings in the Conference North with the only break in form being in the FA Trophy last year at the Martin and Co Arena.
It was a defeat which left Trinity boss Steve Housham disappointed.
Housham said: “I’m very disappointed. We’ve not turned up for most of the game and it took them a red card for them to get us in the game.”
“In possession against the ten we couldn’t keep the ball and we worked in training all week on crossing the ball and we have had four in 90 minutes.”
“We set up how we wanted to come and do things and it didn’t work. It looked like we had been playing Tuesday, Wednesday, Thursday, Friday even though they were the ones that had the game on Tuesday, we looked the leggiest out of the lot.”
“We got ourselves back in the game but some of the goals we had given away were mistakes and we got punished for them.”
“Jan holds his hands up for the third goal. He’s called to win the ball and missed it, but you don’t leave your marker and he had a free tap in. Little things like that have killed us and it’s cost us the game today.”
Ahead of kick off, Trinity manager Steve Housham was enforced into a change as centre half Adam Quinn had been suffering with an abdominal strain.
Quinn is expected to go for some tests to see whether or not it could be a hernia and was replaced in the side by last week’s hero Matt Wilson.
The rest of the side that dominated Droylsden in the first half remained unchanged despite the concerns over fitness of Marc Newsham and Jonathan D’Laryea.
Gainsborough started brightly and won two early free kicks in dangerous positions within the opening ten minutes. Both were delivered in by Jamie Yates and met first by Nathan Jarman who headed over the bar and for the second Newsham headed wide.
Despite the chances and possession it was in fact the Saints who got their noses in front and it was from a free kick of their own.
It was Glenn Walker who delivered the cross into former Derby and Rotherham defender Pablo Mills at the near post and he slotted the ball passed Jan Budtz to give Brackley the lead.
The hosts revelled in their advantage and became more confident on the ball and getting into some dangerous positions.
Trinity managed to hold their ground and prevent any further deficit thanks to solid defending.
However, the first half got worse for the Blues as Jamie Yates was forced off with a back injury just after the half hour mark and replaced by Nick Bignall.
The attacking change unbalanced the Gainsborough side and Brackley were unlucky not to go further in front when Steve Diggin drove at the defence, jinked into space and thumped the post with a venomous shot.
The Saints faithful did not have to wait too long to double the lead as five minutes after the break James Akintunde scored on his debut.
The winger cut in and hit a decent strike that looped over Budtz into the net via a deflection.
It all changed ten minutes later as from a Simon Russell corner Brackley striker Richard Batchelor managed to turn the ball into his own net when back defending.
In the aftermath, centre forward Batchelor was given his marching orders from the referee. It was uncertain what for and it could have been for trying to prevent the goal with his hand.
The Blues had the remaining 30 minutes to get something against ten men and was the ten who almost increased their lead as Akintunde managed to hit the post from a counter attack.
Trinity responded in the best fashion possible as from a long ball from the back, Marc Newsham outmuscled his man before calmly dispatching the equaliser.
Liam Davis replaced Danny Forrest to add more impetus into the attack but once more the change had a negative effect.
A free kick was pumped into the box, Trinity keeper Budtz came out to claim, missed it and Mills grabbed his second as he took advantage of the open goal to restore the Saints lead.
Assistant manager Darryn Stamp was thrown on to try and grab an equaliser but it came too no avail as Brackley managed to see out the game.
Blues boss Housham was deflated with the result.
Gainsborough Trinity: Budtz, Roma, Lacey, D’Laryea, Wilson, Picton, Russell, Forrest (Davis 73), Newsham (Stamp 81), Jarman, Yates (Bignall 34). Subs (not used): Binns, Brogan.
Brackley Town: Crane, Hawtin, McDonald, Clarke, Graham, Mills, Odhiambo, Walker, Akintunde, Diggin, Batchelor. Subs (not used): Austin, Moyo, Truelove, Whittall, Morley.
Goalscorers: Own goal 60, Newsham 71; Mills 26, 80, Akintunde 51.
Red card: Batchelor.
Referee: Mr S Barrow. Assistants: Mr R Gould, Mr D Wilding.