Southampton will enter the Under-21 Premier League Cup with a trip to Crystal Palace or Exeter City next month.
Clubs with Category 1 academies begin their involvement in the Round of 32 of the competition, which is in its inaugural year.
There have been two rounds of regional qualifiers for clubs with Category 2 and 3 academies, with Palace due to host Exeter at Selhurst Park next week.
The winners of that tie will then host Saints' Development Squad in the Round of 32, which is due to be played within 14 days of Monday 18th November 2013.
The Round of 16 will be the first stage at which the Under-21 Premier League Cup will be contested on a national basis. The final will be played over two legs, which are home and away at the main stadia of both teams.