Conference North side Stockport County will host the Blues in the FA Cup fourth qualifying round.
Steve Burr's side will travel to Edgeley Park on Saturday October 25 with the winners progressing to the first round proper.
Chester have not faced the Hatters since a goalless draw in April 2008 when both clubs were still in League Two.
Stockport were relegated from the Football League after 106 years in 2011 and dropped again at the end of the 2012-13 season.
Alan Lord's side are sixth in the Conference North this season and a crowd of 2,562 watch them beat Ilkeston 1-0 in the previous round on Saturday.
Chester will take a good following to Edgeley Park and the tie has the potential to attract the biggest crowd of the round.
