Taking the bus is the cheapest way to travel from Rome to Florence. Buses traveling from Rome to Florence depart from Rome's Tiburtina bus station and take approximately 3 hours to arrive at Florence's Piazzale Montelungo bus station. Approximately 20 buses travel from Rome to Florence every day, with the first bus departing Rome at 1:30 a.m. Bus companies that travel from Rome to Florence include Baltour and BusCenter.

Taking the train is the fastest way to travel from Rome to Florence. Trains traveling from Rome to Florence depart from either Rome's Termini, Tiburtina, or Ostiense train station and take just over an hour to arrive at either Florence's Rifredi, Campo di Marte, or Santa Maria Novella train station. Over 80 trains travel from Rome to Florence every day, with the earliest train departing Rome at 5:53 a.m. Train companies that travel from Rome to Florence include both Trenitalia and Italo.

Taking the train is the most popular way to travel from Rome to Florence. Traveling from Rome to Florence by train is nearly 2 hours faster than taking the bus. Trains are also only €12 more expensive than buses on average. Rome and Florence are located too close together to make flying a prudent option. Buses remain an attractive option for people who would prefer to save while traveling from Rome to Florence.