Ayers started recording as a bebop sideman in 1962 and rose to prominence when he dropped out of City College and joined jazz flautist Herbie Mann in 1966.
In the early 1970s, Roy Ayers started his own band called Roy Ayers Ubiquity, a name he chose because ubiquity means a state of being everywhere at the same time.
Ayers was responsible for the highly regarded soundtrack to Jack Hill's 1973 blaxploitation film Coffy, which starred Pam Grier. He later moved from a jazz-funk sound to R&B, as seen on Mystic Voyage, which featured the songs "Evolution" and the underground disco hit "Brother Green (The Disco King)", as well as the title track from his 1976 album Everybody Loves the Sunshine.
In 1977, Ayers produced an album by the group RAMP, Come into Knowledge. That fall, he had his biggest hit with "Running Away".
In late 1979, Ayers scored his only top ten single on Billboard's Hot Disco/Dance chart with "Don't Stop The Feeling", which was also the leadoff single from his 1980 album No Stranger to Love, whose title track was sampled in Jill Scott's 2001 song "Watching Me" from her debut album Who Is Jill Scott?
In the late 1970s, Ayers toured in Nigeria for six weeks with Afrobeat pioneer Fela Kuti, one of the African continent's most recognizable musicians. In 1980, Phonodisk released Music of Many Colors in Nigeria, featuring one side led by Ayers' group and the other led by Africa '70.
In 1981, Ayers produced an album with the singer Sylvia Striplin, Give Me Your Love (Uno Melodic Records, 1981. That same year, he also produced a second album called Africa, Center of the World on Polydor records along with James Bedford and Ayers's bass player William Henry Allen. Allen can be heard talking to his daughter on the track "Intro/The River Niger". The album was recorded at the Sigma Sound Studios, New York.
Ayers performed a solo on John "Jellybean" Benitez's production of Whitney Houston's "Love Will Save The Day" from her second multi-Platinum studio album Whitney. The single was released in July 1988 by Arista Records.
Ayers has played his live act for millions of people across the globe, including Japan, Australia, England and other parts of Europe.
Ayers is known for helping to popularize feel good music in the 1970s, stating that "I like that happy feeling all of the time, so that ingredient is still there. I try to generate that because it's the natural way I am". The types of music that he used to do this consisted of funk, salsa, jazz, rock, soul and rap.