This is the collection of Sanger Tree of Life Wet Laboratory protocols used for the generation of reference level genome assemblies. This collection currently contains protocols which cover all stages of the wet laboratory processes performed by the Sanger Tree of Life; these processes are sample preparation, sample homogenisation, HMW DNA extraction, HMW DNA fragmentation, fragmented DNA clean-up and RNA extraction. This collection will be updated as protocols within the Tree of Life Core Laboratory continue to be developed and refined.