A late goal from Ben Strevens denied Tranmere a point as they fell to a 2-1 defeat at Brentford.
Joss Labadie's second half strike looked set to earn Rovers a 1-1 draw after they fell behind to a deflection, but substitute Streven scored with just four minutes remaining.
The result was harsh on Les Parry's side who were on top for much of the first half and created plenty of chances throughout.
There was one change to the side that started against Exeter with the injured Paul McLaren replaced by Gareth Edds in midfield.
Rovers enjoyed a decent start to the game with Joss Labadie and Ian Thomas-Moore showing some neat touches and the later forcing home keeper Wojicech Szczesny to palm over his bar on four minutes.
Labadie volleyed just over the bar from long range on six minutes while Ian Goodison did well to clear at the other end four minutes later as Brentford quickly broke into the Tranmere half.
Good work from Craig Curran and Thomas-Moore gave Labadie another chance on 11 minutes and his effort again went narrowly over the top.
Bas Savage's low shot from the edge of the area deflected past the post as Rovers again pressed forward but Brentford remained a threat and Karleigh Osborne fired just over Peter Gulacsi's bar on 30 minutes.
Curran sent an effort from outside the area straight at Szczesny on 31 minutes, Thomas-Moore had a shot blocked and Edds headed at the keeper three minutes before half time.
After doing well in the first half, Rovers had the worst possible start to the second. A shot from Kevin O'Connor looked set to be saved by Gulacsi but the ball took a deflection off a Tranmere player and left the keeper with no chance.
A Thomas-Moore header from Labadie's corner went straight at Szczesny as Rovers looked for a quick response.
Tranmere's next shot did bring them level. A free kick taken by Labadie hit the wall, but the ball rebounded straight to the midfielder and he rifled it straight into the net for his second goal in four days.
Sam Wood fired wide for the hosts on 69 minutes following a free kick and a yellow card awarded to John Welsh while Gulacsi did well to save from Toumanai Diagouraga four minutes later though the shot would have been ruled out for offside had it gone in.
Andy Robinson replaced Savage with 15 minutes remaining, the striker coming on for his second appearance since joining on loan from Leeds.
Six minutes from time Robinson volleyed just over the bar but moments later the home side scored again.
After some pressure from Tranmere deep in the Brentford half, the Bees broke through Charlie MacDonald whose low cross was turned in by substitute Ben Strevens.
Rovers responded by bringing on Kithson Bain in place of Edds while Labadie forced Szczesny to save at his near post with two minutes remaining.
Four minutes of added time gave Tranmere late hope but the closest they came was when Marlon Broomes saw a header turned over by the keeper.
Brentford: Szczesny, Foster, Osborne, Legge, Wood, Grabban (Ainsworth 85), O'Connor, Diagouraga, Weston, MacDonald, Cort (Strevens 44). Substitutes not used: Bull, Blake, Saunders, Hunt.
Tranmere: Gulacsi, Broomes, Taylor, Goodison, Bakayogo, Labadie, Edds (Bain 88), Welsh, Curran, Thomas-Moore, Savage (Robinson 75). Substitutes not used: Coughlin, Gordon, Mahon, Barnett, Gornell.
Referee: David Phillips
Yellow cards: Osborne (62), Welsh (69), Thomas-Moore (82)