Life has evolved from a single origin to generate >1.5 million eukaryotic species. Sequencing all species will provide an inventory of life, transform understanding of evolution, catalogue eukaryotic gene toolkits for biology and biotechnology, and enable monitoring of ecosystems under increasing stress. The Darwin Tree of Life (DToL) is a new initiative that will exploit long read technologies to sequence all 60000 species in the British Isles and play a leading role in the Earth BioGenome Project. This data resource will underpin bioscience for the coming century. We are a consortium of partners who will build and prove an end-to-end pipeline of sample collection, sequencing, genome assembly, annotation and data dissemination that can deliver this visionary project. We will: - Establish sample collection networks (to collect, record and voucher ~8000 species) - Put in place large-scale sequencing and analytic processes (including for single cells and small-bodied taxa) - Generate reference quality, deeply annotated genome assemblies for 2000 species - Develop portals to disseminate the reference genomes, empowering wider scientific communities to embrace genomics in their future endeavours - Share expertise in protocol development and informatics among the Darwin Tree of Life partners to strengthen institutional capacities across the consortium, and with the global EBP.