2019/08/27 | Dolni Benesov | | 2-3 | | SK Prostejov | | | | |
| Slovan Rosice | | 1-5 | | Sigma Olomouc | | | | |
| Valasské Mezirici | | 0-5 | | Trinec | | | | |
| SFK Vrchovina | | 0-5 | | 1. FC Slovácko | | | | |
2019/08/28 | Olešnice U Bouzova | | 0-6 | | SFC Opava | | | | |
| Marianske Lazne | | 2-5 | | Slovan Liberec | | | | |
| Sokol Lanzhot | | 1-3 | | Baník Ostrava | | | | |
| FC Písek | | 1-3 | | Teplice | | | | |
| SK Zápy | | 2-3 | | Bohemians 1905 | | | | |
| TJ Stechovice | | 1-4 | | Banik Sokolov | | | | |
| MFK Frýdek-Místek | | 1-3 | | MFK Karviná | | | | |
| Jiskra Usti Orl | | 0-2 | | Ceské Budéjovice | | | | |
| SK Unicov | | 1-4 | | FC Fastav Zlín | | | | |
| Chlumec Nad Cidlinou | | 2-1 | | 1.FK Pribram | | | | |
| Táborsko | | 4-4 (3-5 Pen.) | | FK Varnsdorf | | | | |
| Kraluv Dvur | | 0-2 | | Dukla Praha | | | | |
| Jiskra Domazlice | | 2-2 (4-3 Pen.) | | FK Ústí nad Labem | | | | |
| Odra Petrkovice | | 3-4 Pro. | | Zbrojovka Brno | | | | |
| Motorlet Prague | | 0-5 | | FK Slavoj Vysehrad | | | | |
| Zdirec Nad Doubravou | | 0-4 | | SK Lisen | | | | |
| FK Prepere | | 0-1 | | MFK Chrudim | | | | |
| FK Loko Vltavín | | 2-1 | | FC Sellier & Bellot Vlasim | | | | |
| Slovan Velvary | | 0-1 | | Viktoria Zizkov | | | | |
2019/09/03 | SK Benesov | | 2-1 | | FK Pardubice | | | | |
2019/09/04 | FK Admira | | 1-2 | | FC Hradec Králové | | | | |
| Sokol Hostoun | | 1-6 | | Jihlava | | | | |
2019/09/10 | FC Hlucin | | 4-2 | | FC Vítkovice | | | | |