2013/05/22 | Minnesota United | | 0-1 | | Des Moines Menace | | | | |
| Rochester New York | | 1-0 | | Portland Phoenix | | | | |
| Ocean City Nor´easters | | 1-0 | | Pittsburgh Riverhounds | | | | |
| Austin Aztex | | 0-2 | | Wilmington Hammerheads | | | | |
| San Antonio Scorpions | | 2-2 (3-4 Pen.) | | FC Tucson | | | | |
| Portland Timbers S23 | | 0-1 | | Charleston Battery | | | | |
| Los Angeles Blues | | 5-1 | | Ventura Fusion | | | | |
| Fort Lauderdale Strikers | | 1-1 (7-6 Pen.) | | Laredo Heat | | | | |
| Tampa Flames | | 1-2 | | Tampa Bay Rowdies | | | | |
| Carolina RailHawks | | 3-1 | | Carolina Dynamo | | | | |
| Richmond Kickers | | 4-1 | | Icon FC | | | | |
| Ocala Stampede | | 1-2 | | Orlando City | | | | |
| Charlotte Eagles | | 3-0 | | Seattle Sounders S23 | | | | |
| Dayton Dutch Lions | | 4-1 Pro. | | Dearborn Stars | | | | |
| Georgia Revolution | | 2-3 | | Atlanta Silverbacks | | | | |
| Reading United | | 1-0 | | Harrisburg City Islanders | | | | |