Tranmere fell to a 2-0 defeat against Cheltenham Town at Whaddon Road. A Danny Holmes own goal on seven minutes was followed by a second half strike by Troy Brown to give the Gloucestershire side the points.
Rovers manager Micky Adams made three changes to the side which started at Portsmouth on Tuesday evening. Defenders Rob Taylor and Josh Thompson missed out through injuries picked up at Fratton Park while Marc Laird dropped down to the bench. Michael Ihiekwe and Lee Molyneux came back into the side while there was a debut for striker Jordan Hugill who joined on loan from Preston North End this week.
Tranmere had the first chance of the game on four minutes when a free-kick from the right was whipped in by Lee Molyneux and the ball landed at the feet of Kayode Odejayi whose shot was saved by Cheltenham goalkeeper Trevor Carson.
Cheltenham took the lead on seven minutes. Shaun Harrad drilled in a low cross from the right and the ball deflected in off Danny Holmes past Owain Fon Williams.
Harrad had a chance minutes later when he picked the ball up in a similar position on the right but his shot was well saved by Fon Williams while on 20 minutes, Wes Burns picked the ball up on the right and sent a shot over the crossbar.
On 32 minutes, Lee Molyneux had a cross stopped by Carson before moments later a shot by Jennison Myrie-Williams from the edge of the penalty area flew just over the crossbar.
Five minutes later, Myrie-Williams picked up the ball on the left side of the box and drilled a low shot which was blocked away for a corner with Max Power's set piece deflected just over the crossbar only for the referee to award a foul on a Cheltenham defender.
Rory Donnelly had the final opportunity of the first 45 minutes as he struck a bobbling ball from 30 yards but his effort drifted wide of the target as Cheltenham went in ahead at the break.
Three minutes after the restart, Jordan Hugill nipped in ahead of Carson to intercept a back pass but his shot was cleared off the line by Cheltenham defender Troy Brown.
Cheltenham doubled their lead on 61 minutes. A deep free-kick from the left by Matt Richards was met at the back post by Troy Brown whose effort went back across the goal and into the net.
Rovers manager Micky Adams then made a double substitution with Marc Laird and Iain Hume coming on in place of Rory Donnelly and Kayode Odejayi.
Hugill had a shot blocked following a Molyneux free-kick before Tranmere made their final change of the game on 78 minutes with Jake Kirby introduced in place of Molyneux.
With nine minutes remaining, Power played the ball out to the right where Hume laid it inside to Hugill whose goalbound shot was well saved by Carson.
On 87 minutes, Laird's cross from the right found Myrie-Williams at the back post but his header bounced back out off the post before the Cheltenham defence scrambled clear while a Hume snapshot was blocked on its way to goal but it was Cheltenham who held on to take all three points.
Cheltenham: Carson, Vaughan, Taylor, Brown, Braham-Barrett, Burns (Berry 88), Mills, Sparrow (Hanks 90+4), Matt Richards, Eliot Richards, Harrad (Johnstone 78). Substitutes: Reynolds, Kotwica, Wynter, Manset.
Tranmere: Fon Williams, Holmes, Dugdale, Ihiekwe, Molyneux (Kirby 78), Myrie-Williams, Jennings, Power, Donnelly (Laird 62), Hugill, Odejayi (Hume 62). Substitutes: Pilling, McGahey, Ridehalgh, Koumas.
Referee: Michael Bull
Yellow cards: Myrie-Williams (20), Odejayi (28), Holmes (59), Brown (64), Burns (66)