Tranmere's 18 match unbeaten home run came to an end with a 2-1 defeat against Doncaster Rovers.
Ronnie Moore's side had by far the majority of the game's chances but conceded early in the second half.
It looked like they'd gained a share of the points when Jake Cassidy scored his ninth goal of the season on 86 minutes but former Rovers striker Iain Hume won the game for Doncaster in the 92nd minute.
Despite the result, Tranmere's four point gap at the start of play meant they still led League 1 at the final whistle.
Ronnie Moore made one change to the team that started at Bournemouth with Andy Robinson returning from injury to take the place of the injured Jake Jervis while Ian Goodison was named on the substitute for the first time in 2012-13.
Robinson almost opened the scoring in the fifth minute with a swerving drive from 20 yards but his right foot shot drifted inches away from the top right corner of Gary Woods' goal.
Doncaster's Billy Paynter forced a save out of Owain Fon Williams with a back post header on five minutes while a free kick from Robinson at the other end was curled straight at Woods from the edge of the area.
Danny Holmes sent a dipping volley wide of the target from 30 yards on 13 minutes and Robinson missed from a similar distance two minutes later after receiving a pass from James Wallace.
A Ben Gibson header missed Woods' goal on 17 minutes and Rovers kept the pressure up but Adam McGurk couldn't control a wonderful cross from Zoumana Bakayogo at the far post.
The visitors couldn't have gone much closer to breaking the deadlock on 25 minutes. A left wing cross was met by Paynter at the near post and his header hit the underside of the crossbar and bounced on the line and away from goal.
McGurk couldn't quite intercept a short header back to goal by Tommy Spurr on 28 minutes and Wallace sent a close range header over the bar from McGurk's corner four minutes later.
Despite clearly being in some discomfort, Robinson forced a great save out of Woods with a stunning curling shot from from wide on the left on 36 minutes and the midfielder fired inches over the bar three minutes later after running on to Wallaces's pass.
Bakayogo volleyed wide of the near post from McGurk's corner on 44 minutes and the final chance of the half saw McGurk's right foot effort deflect into the side netting.
Doncaster took the lead with the first attack of the second half. A high ball into the Rovers area fell between Ash Taylor and Ben Gibson and Paynter took advantage to get between the pair and fire a low shot past Fon Williams from 12 yards.
McGurk tried to level with a shot after twisting and turning his way past three challenges inside the Doncaster area on 49 minutes but his effort was headed clear while two attempts from Palmer from outside the area failed to hit the target soon after.
After a Robinson shot from 20 yards deflected just past the left hand post on 64 minutes, the former Swansea player's corner was headed a couple of yards wide by Wallace.
Robinson was replaced by Joe Thompson on 70 minutes and Cole Stockton came on for McGurk seven minutes later as Moore freshened up the attacking options.
Doncaster cleared twice from under their own bar on 79 minutes before Thompson shot high over the bar from the left side of the area.
The equaliser finally came on 86 minutes. Wallace and Bakayogo traded passes on the left wing before the former's cross was headed back across goal by Abdulai Bell-Baggie and after, Doncaster couldn't react, fired in by Cassidy at the back post.
However, two minutes into added time Doncaster stole the points, with Hume firing between the legs of Fon Williams from a tight angle.
Rovers threw everyone forward late on and had a couple of penalty appeals turned down but Doncaster left Prenton Park with all three points.
Tranmere: Fon Williams, Holmes, Taylor, Gibson, Bakayogo, Bell-Baggie, Wallace, Palmer, Robinson (Thompson 70), McGurk (Stockton 77), Cassidy. Substitutes: Mooney, Kay, Goodison, Black, Harrison.
Doncaster: Gary Woods, Quinn, Husband (Griffin 77), Jones, Paynter (Blake 90+3), Cotterill (Syers 61), Spurr, Hume, Harper, Bennett, Keegan. Substitutes: Sullivan, Brown, Michael Woods, Wakefield.
Referee: Scott Mathieson
Yellow cards: Wallace (44)