title: genomic DNA of Picochlorum sp. BPE23
design: 50 mg washed and lyophilized biomass was used for gDNA extraction using MagMax Plant DNA kit (Applied Biosystems). Extracted gDNA was sheared using a megadisruptor device (Diagenode), aiming for a 25 kb fragment size. Barcoded SMRT bell template libraries were made for each microalgal strain using SMRT bell express template prep kit 2.0 (PacBio). gDNA was eluted in 20 L elution buffer and pooled for size selection on a Blue Pippin instrument (Saga Sciences). The 8 to 13 kb and >=13 kb fractions were collected for each lane after which purification and concentration was performed with Ampure XP beads (Beckman Coulter). Purified SMRT bells were finally analyzed for size and used for primer annealing, polymerase binding, and purified using AMPure PB beads (SMRTLink10.1). Sequencing was done on one SMRT cell of a PacBio Sequel IIe system with 65 pM on plate loading by diffusion with standard settings and 30 hour movie time. Data demultiplexing and HiFi read processing was done by SMRT Link 10.1.