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Live performances

The a cappella opening was too complex to perform live, so Mercury tried various ways of introducing the song. When " Mustapha " became a live favourite, Mercury would often sub in that song's a cappella opening, which was easier to reproduce live as it was only one voice—this combination features in their 1979 live album Live Killers. During their 1982 Hot Space Tour, and occasionally at other times, Mercury would do a piano improvisation (generally the introduction to "Death on Two Legs") that ended with the first notes of the song. Often, the preceding song would end, and Mercury would sit at the piano, say a quick word, and start playing the ballad section. At Live Aid where "Bohemian Rhapsody" was their opening song, Mercury commenced with the ballad section.

Initially following the song's release, the operatic middle section proved a problem for the band. Because of extensive multi-tracking, it could not be performed on stage. The band did not have enough of a break between the Sheer Heart Attack and A Night at the Opera tours to find a way to make it work live, so they split the song into three sections that were played throughout the night. The opening and closing ballads were played as part of a medley, with " Killer Queen " and "March of the Black Queen" taking the place of the operatic and hard rock sections. In 1976 concerts where the same medley was played, the operatic section from the album would be played from tape as the introduction to the setlist. During this playback, Mercury would appear briefly to sing live for the line, "I see a little sillhouetto of a man". As the song segued into the hard rock section, the band would emerge on the smoke-filled stage—the playback would end at this point, and the hard rock section would be performed live (without the final ballad section, which appeared later in the set).

Starting with the A Day at the Races Tour in 1977, the band adopted their lasting way of playing the song live. The opening ballad would be played on stage, and after May's guitar solo, the lights would go down, the band would leave the stage, and the operatic section would be played from tape, while coloured stage lights provided a light show based around the voices of the opera section. Most playings of the opera section from the tape would often be accompanied by a portion of the song's music video containing the footage used for the operatic portion of the song. Other playings would be played over montages of footage filmed from the band members' other experiences throughout their daily lives. A blast of pyrotechnics after Taylor's high note on the final "for me" would announce the band's return for the hard rock section and closing ballad. Queen played the song in this form all through the Magic Tour of 1986. This style was also used for the Freddie Mercury Tribute Concert, with Elton John singing the opening ballad and then after the taped operatic section, Axl Rose singing the hard rock section. John and Rose sang the closing ballad part together in a duet.

"Bohemian Rhapsody" was performed by Queen + Paul Rodgers throughout their tours, accompanied by a video of Mercury. Footage from the Live at Wembley '86 was used for the 2005–06 tour, and the 1981 Montreal performance used for the Rock the Cosmos Tour. As with the Queen tours, the band went backstage for the operatic section, which was accompanied by a video tribute to Freddie Mercury. When the hard rock section began, the lights came back up to the full band on stage, including Rodgers, who took over lead vocals. Rodgers duetted with the recording of Mercury for the "outro" section, allowing the audience to sing the final "Nothing really matters to me", while the taped Mercury took a bow for the crowd. Rodgers would then repeat the line, and the final line ("Any way the wind blows") was delivered with one last shot of Mercury smiling at the audience. Commenting upon this staging, Brian May says that they "had to rise to the challenge of getting Freddie in there in a way which gave him his rightful place, but without demeaning Paul in any way. It also kept us live and 'present', although conscious and proud of our past, as we logically should be."

Since 2012, May and Taylor have toured with former American Idol contestant Adam Lambert under the name Queen + Adam Lambert (following two one-off performances together in 2009 and 2011), with "Bohemian Rhapsody" regularly included at the end of their set.