Victory against the Czechs at Wembley would see England top Group D with seven points, setting up a last-16 meeting with the runner-up of Group F - which at this stage could potentially be France, Germany or Portugal.
A draw would see the Czech Republic go through as group winners and England advance as runners-up. Southgate’s side would then play the runner-up of Group E, which includes Spain, Poland, Sweden and Slovakia, while defeat could see England finish in third depending on results elsewhere.
Here’s all you need to know ahead of this evening’s match.
When is it?
The match will kick off at 8:00 pm BST on Tuesday 22 June.
How can I watch and will there be a live stream?
The match is live on ITV1 with coverage starting from 7:00pm. It can also be live streamed online via the ITV Hub, click here.
Czech Republic XI: Vaclik; Coufal, Celustka, Kalas, Boril; Soucek, Holes, Masopust, Darida, Jankto; Schick
Substitutes – Koubek, Sevcik, Mateju, Hlozek, Kral, Brabec, Vydra, Mandous, Krmencik, Kaderabek, Barak, Pekhart
England XI: Pickford; Walker, Stones, Maguire, Shaw; Rice, Phillips; Saka, Grealish, Sterling; Kane
Substitutes – Ramsdale, Johnstone, Henderson, Rashford, Trippier, Mings, Coady, Sancho, Calvert-Lewin, White, James, Bellingham
Czech Republic: 13/2
The stalemate with Scotland brought expectations back down to earth and that may be no bad thing in the long run. Qualification is now assured but Southgate still has the opportunity to make changes and experiment. England smashed five unanswered goals past the Czech Republic at Wembley in qualifying, only to lose in Prague. A narrow win feels likeliest. Czech Republic 0-1 England.