In the decades after the bicycle arrived in Australia, cycling competitions became big business. Talented riders including Hubert Opperman and Ken Ross abandoned their amateur status to vie for cash prizes in major competitions in Australia and overseas.

Cycling's popularity declined in the mid-20th century, but underwent a major resurgence from the 1980s, with government funding for high-performance programs.

Australians continue to perform well in the Tour de France, with yellow jersey wearers including Phil Anderson (1981), Stuart O’Grady (1998 and 2001), Bradley McGee (2003), Robbie McEwen (2004), Cadel Evans (2008, 2010 and 2011), Simon Gerrans (2013), Rohan Dennis (2015), Jai Hindley (2023).

Australia's first Olympic cycling medal was won by Dunc Grey and the Museum's collection includes a UCI World Championship medal won by Sue Powell. Sue, a multiple world champion medallist, also won gold in the 3-kilometre individual pursuit (C4 classification) and silver in the road race at the 2012 Paralympics.