Illumina Genome Sequencing of O. tipulae CEW1: L1 larvae
Egg-laying worms were washed off plates and passed through a 70-m mesh. This step allows existing eggs to fall through the mesh, leaving the mature worms on the top. These worms were then bleached with sodium hypochlorite solution (Sigma Cat# 425044-250ML, 0.5 M NaOH, 3% bleach) to collect embryos (largely 2-4 cell stage). Early embryos were further developed in Virgin S Basal (VSB) medium (100 mM NaCl, 5.7 mM K2HPO4, 44.1 mM KH2PO4) at 25C to reach to the desired embryonic stages. To prepare larval stages, the flow-through of the 70-_m mesh was further passed through a 25-_m mesh to separate mixed embryos from various stages of worms. These mixed embryoswere developed in the VSB medium until all embryos reach to L1.