Wanderers will find out on Friday who they will meet in the first knockout round of the Papa John’s Trophy.
The field has been narrowed for Ian Evatt’s side by virtue of the fact the draw – which will take place during Sky Sports’ coverage of the Birmingham City v Sunderland game – is still split in northern and southern halves.
Bolton are also guaranteed a home draw as they finished top of their group with seven points from their three games against one of the other group runners-up.
Derby County and Manchester City Under-21s still have to determine who has won their group – but the second placed side will go into the hat as one of seven teams that the Whites could face including Barrow, Bradford City, Accrington Stanley, Morecambe, Wolves Under-21s and Barnsley.
Evatt has spoken about his disappointment at having been knocked out in the last 16 by League Two Hartlepool United last season and, speaking last month, said he would love to lead Wanderers towards Wembley once again.
“I think that anytime you get a chance to play a cup final at Wembley you should try and take it with both hands because those are the moments in your football career that you remember.
“I was lucky enough on a couple of occasions to play there and they really are the games that stand out above the rest. Your families want to come and see their kids play at Wembley Stadium and hopefully we’ve got a chance of doing that.
“You look at the other cup competitions, who wins the Carabao Cup every season and who wins the FA Cup every season, it’s always one of the big four predominantly and in this competition we’re one of the better teams in League One so we have to try and target it and we will do.
“We think we have got the squad capable of it. It is stronger again from last year and as you can see from tonight, we’re taking it very seriously.”
The exact date of the last 32 fixture has not yet been confirmed but will take place in the week beginning November 21.
Meanwhile, both Dapo Afolayan and George Johnston missed out in their respective categories in the North West Football awards, held on Monday night.
Afolayan was nominated for League One player of the year, with the award eventually going to Will Keane of Wigan Athletic.
Johnston was nominated for the Rising Star award, which eventually went to Everton's Anthony Gordon.
Wanderers did have some success on the night, however, with a win in the Best Club Marketing, Sponsorship or Engagement Campaign.
The 'Be Part of It' season ticket offer proved hugely successful, helping ensure average crowds of more than 18,000 so far at the University of Bolton Stadium this season.
The team was congratulated on social media by club CEO Neil Hart, who said: "Although I could not attend last night, delighted to see the team collect this award. Special mention to Siobhan Chaplow and Daniel Scott for making this happen, and of course the Bolton Wanderers Supporters’ Trust for our fantastic partnership together and working in collaboration we are certainly stronger."
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