[1] The tournament proper will feature both professional and amateur teams in the United States soccer league system.

To be eligible, teams must be members in good standing of their leagues on December 31, 2023, and remain so through the 2024 final.

Beginning in 2024, the United Premier Soccer League Spring Champion also earned a direct entry into the first round proper of the tournament.

The teams will be organize into 14 regional pools prior to the qualifying round draw[1] A total of 25 states and the District of Columbia are represented by clubs in the U.S. Open Cup Qualification this year.

Match ups, byes and home teams in each geographical group were determined by random selection.