Taking the bus is the cheapest way to travel from Rome to Pisa. Buses traveling from Rome to Pisa depart from Rome's Tiburtina bus station and take just over 5 hours to arrive at Pisa's Autostazione Via Pietrasantina. Approximately 6 buses travel from Rome to Pisa every day, with the first bus departing Rome at 2:00 a.m. Bus companies that travel from Rome to Pisa include Baltour and Flixbus. Ridesharing services like BlaBlaCar also frequently offer trips from Rome to Pisa.

Taking the train is the fastest way to travel from Rome to Pisa. Trains traveling from Rome to Pisa depart from either Rome's Termini, Tiburtina, Trastevere, or Ostiense train station and take as little as 2 hours to arrive at Pisa's Centrale train station when traveling on a high-speed train. Over 60 trains travel from Rome to Pisa every day, with the earliest train departing Rome at 6:05 a.m. All trains that travel from Rome to Pisa are operated by Trenitalia with a variety of train types available.

Taking the train is the most popular way to travel from Rome to Pisa. Traveling from Rome to Pisa by high-speed train is approximately 3 hours faster than taking the bus. Trains are also only €16 more expensive than buses on average. Rome and Pisa 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 Pisa.