The Three Lions are in Prague to face this summer’s Euro Finals host nation and the head coach is keen to see as many of his players in action as possible in both this game and Monday’s clash with Germany.
Czech Republic v England
U21 International Match
7.30pm, Friday 27 March 2015
Letna Stadium, Prague
Live on fotbal.cz
And with Chambers and Targett in his squad for the first time and only one previous call-up for Bettinelli, Southgate is ready to assess his options ahead of England's return here in June.
"We’ll have a couple of players making their debuts, so we’re looking forward to seeing how they go," he revealed.
"With Jack Robinson now out for the summer and Luke Shaw out injured, it’s a chance for Matty Targett and to see how he copes with that.
"And Marcus hasn’t had a game for us yet either so it's an opportunity."
As Euro hosts, the Czechs have not had to come through the qualifying process and play-offs like the seven other finalists involved in the finals.
But with home support behind them, both on Friday and in the summer, Southgate sees this game as another important test for his side before they return to England ahead of the game with Germany in Middlesbrough.
"Out of the two games that we’ve got, we think Germany will have the mentality to come and play and the Czechs have tended to be more of a counter-attacking team. They’re quite strong, good on the break and good from set-plays," explained the former Aston Villa and Crystal Palace man.
"I think we’ll probably have more of the ball, but they’re still a step up from what we played in our [qualifying] group and they’ve got players like [Watford's Matej] Vydra and [Middlesbrough's Tomas] Kalas who we know from playing in England.
"So it’s a really good test to come and play them on their home turf because the tournament here is picking up momentum."
England have been in Czech Republic since Monday, when they arrived to train 150 miles south-east of Prague in Olomouc.
That will be where the squad are based during the Euro Finals, so this game arrived at a good time for Southgate and his team to take the chance of an early bedding-in period ahead of what will hopefully be a successful summer.
"For us, the whole week has been a good opportunity," he added.
"We’ve stayed at the base camp to get a feel for the place and we’re really pleased with it.
"The players have got a feel for what it’s like and we’re happy with that."
Friday’s game will be streamed live on the Czech FA website here. The match kicks-off at 7.30pm (GMT).