The Singapore Botanic Gardens Heritage Museum is located in Holttum Hall, which is next to the Botany Centre in the Tanglin Core of the Gardens. Built-in 1921 to serve as the Director of the Gardens' office and laboratory, the historic two-story building has been designated as an Urban Redevelopment Authority conservation building. This was where Professor Eric Holttum, the Gardens' Director from 1925 to 1949, developed the orchid breeding technique which enabled the efficient production of healthy seedlings in sterile flasks from hybrid seed. This technique, which is still used today, was a key factor that stimulated orchid industries throughout the region.

The museum features many hands-on interactives, multimedia films, and period-inspired illustrative graphics that detail the Gardens' rich heritage and significance in botanical research, historical growth, and connection to the nation’s distinctive history. Historical objects that are on display include rare photographs, artifacts, plant specimens, rare botanical books, and botanical paintings that date back to the early 19th century. The completion and opening of this historic museum helped earn the Singapore Botanic Gardens as Singapore’s first UNESCO World Heritage Site in 2015.