Mount made two substitute appearances for England last month, coming on in the second half against Bulgaria and Kosovo.
The midfielder has scored twice in four Premier League games for Chelsea since then and his performances have convinced Gareth Southgate he is ready to start for the Three Lions.
Chelsea manager Frank Lampard has repeatedly selected Mount ahead of Ross Barkley this season, and it is believed the former Everton midfielder will again make way for Mount in Prague.
England have won all four of their qualifiers for Euro 2020 and will guarantee their place at the tournament with a victory on Friday.
The Three Lions then travel to face Bulgaria on Monday.
England will be without midfielder James Maddison, who has been pulled out of the squad through illness.
Maddison has been an unused substitute four times in two different England squads but is yet to receive his first senior cap.
Asked last week why the 22-year-old has not featured yet, Southgate said: “I can’t guarantee people football.
“He only has to see the players that have been left out to recognise just to get in the squad is an achievement, and I don’t handle him any differently to any of the others, really.
“He wants to talk football which is great. He wants to know what areas of his game that we like and that we think he can improve.
“He seems to take all of that on board really well, so he’s playing very well, but so is [Mason] Mount, so is [Jadon] Sancho and so are all of the others – that’s the conundrum.”
Southgate believes he now has enough attacking talent at his disposal to not have to rely on one or two key players at next summer’s tournament.
“We’ve got Callum Hudson-Odoi coming through and other players are emerging so quickly,” he said.
“You’ve got Bukayo Saka at Arsenal just dipping his toes in the first team as well. So, we’ve got really good players, we need more than three.
“I’ve always said that in tournaments that I played in, we always pinned our hopes on one player or two players.
“Then there was an injury, a drama and a crisis and you need players to be able to start games and you need players who are able to come on and make a difference in games.
“So, I don’t think it matters what our starting three is. Clearly, at the moment Raheem [Sterling] and Harry [Kane], their numbers are incredible and the consistency of their performance is, but the rest can be down to performance at any given time.
“And how we see the guys in training and their physical level. These guys know that Tammy coming through is going to push that.
“Callum Wilson is going to want to push – we can try to motivate the players but that competition for places is as much as a driver than anything else.”