Mark Hughes' side put together a four-match unbeaten run after losing to Liverpool on the opening day of the Premier League season, including a 3-1 success over Walsall in the second round of the League Cup.
But that run came to an end at the Emirates Stadium on Sunday, when Mesut Ozil inspired Arsenal to an impressive victory.
Stoke and Tranmere have only met 18 times in their histories, and this will be the first time they have clashed in the League Cup.
Their last encounter came in the third tier of English football in 2001, which ended 2-2, and Tranmere can take heart from the fact they have not lost to Stoke since December 1996.
The last time Stoke won at Prenton Park was two years prior to that victory, and although there is now a significant gulf between the clubs, USA international Geoff Cameron insists the Premier League outfit are fully focused on defeating their League One opponents.
"We won't take Tranmere lightly. It's an important competition for us and it is a chance to keep our confidence going with a win away on Wednesday," he told the club's official website.
Tranmere have knocked out Mansfield and Bolton to get to this stage, but they are currently on a run of four games without a win. Their last match saw them fell to a 2-0 defeat to Notts County on Saturday.
Hughes will be without Spanish defender Marc Muniesa (groin) for the tie, but otherwise has a full squad to select from.
He is expected to give the likes of Wilson Palacios and Stephen Ireland much needed game time, while new signing Marko Arnautovic may start after impressing in his first start at the Emirates.
Tranmere will rest 40-year-old defender Ian Goodison for clash, while Jason Koumas, Andy Robinson, James Wallace and Jake Kirby will all be unavailable through injury.
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