Tranmere Rovers travel to League 2 Bury tonight after they were given a second chance when Accrington Stanley resigned from the competition.
Rovers were knocked out in the first round, on penalties by Stanley, but were handed a reprieve after the League 2 side fielded an ineligible player.
They beat Stockport County in the last round and are now looking to go all the way, as Manager Les Parry told the Liverpool Daily Post: "It is important we beat Bury. I said from round one that this is a competition we are taking seriously.
"We are out of the League Cup and FA Cup now but we knew before the season started that the JPT was our only realistic chance of winning a knockout competition.
"We have players here who are coming to the end of their careers and they've never played at Wembley. We have players who are just starting out their careers and they've not played at Wembley. It is a massive opportunity for them."
Bury are flying high in League 2 having lost just one of their last nine league matches, that coming against Bradford City just over a week ago. They currently sit third in the table with 28 points, that's 12 more than tonight's opponents.
There's a big Yorkshire derby in the Northern Section as Rotherham United host Huddersfield Town.
The Terriers currently sit second in the npower League 1 table after winning eight of their first 15 matches and the Town Manager Lee Clark is eyeing another victory, as he told the Yorkshire Post: "It is a huge game because we are just four ties away from Wembley and I can tell you that I, for one, am taking the competition very seriously.
"I will be fielding the strongest team that is available on the night. There is no chance of any experimenting and there will be no weakening of the team for what is an important match.
"We are going to Rotherham United to win the game."
Rotherham have picked up only two points less from their first 15 matches, with 24 on the board already. But they go in search of their first home win since September 25th, when they beat Chesterfield.
Since then they've played out four draws and suffered on defeat, so who better to beat to get back on track than their local rivals.
There's also a derby in Devon as Plymouth Argyle host Exeter City. The two League 1 clubs have been in indifferent form in recent weeks and both sit further down the table than they would like.
Argyle currently sit 15th with Exeter just one place below them, level on 19 points but a derby match could be just what both sides need, to kick start their season.
Plymouth boss Peter Reid was left disappointed by his side's 4-0 FA Cup defeat to Swindon Town at the weekend but he's expecting more tonight, as he told www.pafc.co.uk: "It shows you how important this game is by what we think the size of the crowd is going to be. It is a big game for us.
"Exeter have had a couple of iffy results recently, so they'll want to bounce back. I should imagine it will be a fast and furious game.
"It was difficult on Saturday, but in the two home games before then we played really well, put up good performances. Hopefully we can do that again."
Exeter also crashed out of the FA Cup at the weekend, as they lost to League 2 Bury and it was the Grecians' fourth consecutive defeat and now Manager Paul Tisdale is hoping for a swift turnaround:
"We have to remain confident," he told the Express and Echo. " A lot's changed in two weeks but we have to remember that we are a good side - it's just been a glum past fortnight.
"The danger is to look too deeply into things and lose faith with what you're doing or lose faith in the players. We can't become anxious as a team. A challenge like this, with a succession of defeats in a short space of time in games away from home, you need to stay strong in your own mind.
"Hopefully it's a blip. It's a rut we have got ourselves into and we have got to respond."
Bristol Rovers Manager Paul Trollope is hoping his side can avoid the second cup shock of the week when they travel to Wycombe Wanderers.
The Pirates were beaten at the weekend by non-league Darlington and Trollope is aware of the potential banana skin they face tonight, as he told www.bristolrovers.co.uk:
"We are disappointed and frustrated, it was a poor result, there is no hiding from that, and we have got to bounce back from that.
"The next game is a Johnstone's Paint Trophy game, we have already stated that we are taking the competition very seriously, and that will definitely be the case.
"The spirit is still good, the belief is still there, it has taken a couple of knocks, but we have to make sure we stick together, stay focused and don't panic.
"We must make sure we produce a performance with the intensity levels and the quality we know we are capable of. If we can do that tomorrow we are confident we can win the game."
Wycombe have lost just one of their last 11 matches in all competitions, that only coming at the hands of league leaders Chesterfield. They currently sit fifth in the table, as they look to bounce back to League 1 at the first attempt.
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