The Italians are on the cusp of booking their spot in Brazil next year, with a victory over the Czechs in Turin set to guarantee their spot with two games still to play.
After seeing off Bulgaria 1-0 on Friday Italy are in a confident mood and Prandelli hopes they can build on that against a Czech side still in the hunt for a play-off spot in UEFA's Group B.
"We have worked very well in the last few days...we have had the time to recover," Prandelli said on Monday.
"We know that this match is going to be very tough and we shouldn't make any mistakes from a mental point of view. We suffered a bit in the match against Bulgaria.
"We know that if we manage to play in a structured way, we can limit the difficulties we have, whereas if we're both tired and disorganized, we can have difficulties against any team.
"(Tuesday) is going to be particularly tough as they will do their best in order to gain a better position in the group.
"We have a very high objective and we want to seal our qualification tomorrow."
Prandelli has yet to decide the make-up of his side against the Czechs, with Milan skipper Riccardo Montolivo a chance to play after missing the Bulgaria victory through suspension.
Montolivo's return could see Prandelli tinker with his formation against the Czech Republic.
Asked about his chances of playing, Montolivo said: "I'm feeling well."
"I've had some problems after the match between Milan and Cagliari but after three or four days it was better and I'm doing well from that point of view.
"As far as the game plan is concerned, there aren't many differences as the midfield made of three players has always been adopted in the last three years, therefore it's the same to play with a defence made of three or four players.
"We're ready to play with different systems. We have already done it during the qualification matches, so we're relaxed from that point of view."
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