Rovers were beaten 2-1 at home against Welling United. After taking the lead through Lee Vaughan, an own goal and Kadell Daniel strike gave Welling three points.
Tranmere were unchanged from the team which beat Kidderminster Harriers 2-0 on Bank Holiday Monday.
Rovers put a lot of pressure on the Welling goal in the early stages and Gary Taylor-Fletcher played a low cross which was met by Jake Kirby at the far post but a combination of goalkeeper Marek Rodak and defender Ian Gayle cleared their lines.
Tranmere took the lead on 18 minutes when Taylor-Fletcher played a lovely dinked pass to Lee Vaughan who volley home past Rodak.
Welling got themselves back in the game almost immediately as a free-kick wasn't cleared and Gayle poke towards goal hit Vaughan and trickled into Scott Davies' goal.
Rovers had a good chance on 25 minutes when Jeff Hughes' free kick from the right was flicked on by James Norwood and Michael Ihiekwe met it but his effort lacked power and Rodak saved.
Davies had to be alert to collect another deflected free kick on 38 minutes before Taylor-Fletcher played in Norwood whose shot was saved low down by Rodak.
Two minutes into the second half, Hughes nodded the ball goalwards only for the ball to be cleared off the line by Welling skipper Barney William as Tranmere looked to retake the lead.
Taylor-Fletcher then sent a 20 yard free kick over the crossbar before Hughes was replaced by Jack Mackreth after taking a knock in midfield.
Steven Jennings had a shot deflected high into the air which was tipped over the bar by Rodak while Scott Fenwick came on for Taylor-Fletcher on 61 minutes.
Then Mackreth made a good run on the right and pulled the ball back to Norwood who turned and shot towards goal only for his effort to be blocked and cleared away.
Welling took the lead on 71 minutes as the broke through the Rovers defence and the ball was rolled to Kadell Daniel who rolled the ball home.
Gary Brabin made his final change of the game by bringing Michael Higdon into the action in place of Kirby on 76 minutes.
With a minute remaining, Norwood hit the post following a deep left wing corner before Higdon fired the rebound just wide as Rovers couldn't find their way back into the game.
Tranmere: Davies, Vaughan, Ihiekwe, McNulty, Hill, Harris, Hughes, Jennings, Kirby (Higdon 76), Taylor-Fletcher (Fenwick 61), Norwood. Substitutes: Turner, Riley.
Welling: Rodak, Chambers, Kashket (Porter 90+2), Nortey, Gayle, Taylor (Wanadio 67), Lokko, Corne, Harris, Williams, Daniel. Substitutes: McIntegart, Saggers, Yusuff.
Referee: Adam Bromley
Yellow cards: Chambers (51)
Goals: Vaughan (18); Vaughan (og 19), Daniel (71)
Attendance: 5173 (63)