this is in the wrong place but I’ll bite anyway.

Flying in such a situation is almost always easier, and this decision usually depends on cost. If you don’t have a railpass, such a long point-to-point ticket is going to cost you but probably still less than flying. However a night train like this could take up to 16 hours and even with a couchette a good nights sleep is difficult leaving you tired the next day. So I’d say the decision depends on how cheap the flight is or whichever you value more, time or money?

It seems like a plane ticket from Rome to Prague is only 40euro. So that reasonable enough and I think it will end up being cheaper and easier to fly. Since I will alone for that part of the trip, as a girl, I don’t really feel too comfortable traveling alone on a night train.

Thanks for the input.

It’s easy to get from Florence to Rome, right?

Very easy. Trains run all the time between the two cities.
http://www.trenitali…
Obviously you’ll still have to get to the airport. Given that Italian trains can be a bit behind schedule, allow yourself plenty of time. And make sure there won’t be a strike on the day you go (or consider going to Rome the night before).

Assuming your flight is from Rome Ciampino, it’s easy to catch a shuttle bus at Termini that will take you there. http://www.terravisi…
But I’d recommend taking an earlier bus than they suggest, because when I took it I was stuck in a massive traffic jam.