title: PacBio HiFi sequencing data for Panulirus interruptus (Revio)
design: High molecular weight (HMW) genomic DNA (gDNA) was extracted from 70 mg of gill tissue using the Nanobind Tissue Big DNA kit according to the manufacturer's instructions (Pacific BioSciences, CA). The DNA purity was estimated using absorbance ratios (260/280 = 1.87 and 260/230 = 2.30) measured on the NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, MA). The final DNA yield (50g) was quantified using the Quantus Fluorometer (QuantiFluor ONE dsDNA Dye assay; Promega, Madison, WI). The size distribution of the HMW DNA was estimated using the Femto Pulse system (Genomic DNA 165 kb kit, Agilent, CA), and found that 84 % of the fragments were 100 kb or longer. The HiFi SMRTbell library was constructed using the SMRTbell prep kit 3.0 (Pacific Biosciences, Menlo Park, CA; Cat. #102-182-700) according to the manufacturer's instructions. HMW gDNA was sheared to a target DNA size distribution between 15-18 kb using Diagenodes Megaruptor 3 system (Diagenode, Belgium; cat. B06010003). The sheared gDNA was concentrated using 1X of SMRTbell cleanup beads provided in the SMRTbell prep kit 3.0 for the repair and a-tailing incubation at 37C for 30 minutes and 65C for 5 minutes, followed by ligation of overhang adapters at 20C for 30 minutes, clean-up using 1X SMRTbell cleanup beads, and nuclease treatment at 37C for 15 minutes. The SMRTbell library was size selected using 3.1X of 35% v/v diluted AMPure PB beads (Pacific Biosciences, Menlo Park, CA; Cat. #100-265-900) to progressively remove SMRTbell templates <5kb. The 15  18 kb average HiFi SMRTbell library was sequenced at UC Davis DNA Technologies Core (Davis, CA) using six 8M SMRT cells (Sequel II sequencing chemistry 2.0 and 30-hour movies - 5 on a PacBio Sequel IIe sequencer and one on a PacBio Sequel II sequencer) and two 25M Revio SMRT cells (Revio sequencing chemistry and 24-hour movies on a PacBio Revio sequencer).