PacBio HiFi whole-genome sequencing of Chrysina woodi
Highmolecular-weight genomic DNA was extracted from thoracic tissue of a single adult Chrysina woodi using the PacBio Nanobind Tissue Big DNA Kit according to the manufacturers protocol. DNA concentration was quantified using the Qubit fluorometric assay, and fragment size distribution was assessed with the Agilent Femto Pulse system. Approximately 4 g of genomic DNA per sample was sheared using a Covaris g-TUBE at 3,200 rpm for 1 minute. Size selection was performed on the Sage Science BluePippin system using an E2 marker and the Range + T protocol, with fragments 11 kb collected using a +20-minute elution to enrich large DNA fragments. SMRTbell whole-genome libraries were prepared using the PacBio SMRTbell Prep Kit 3.0. Polymerase binding was carried out with the PacBio Revio SPRQ Polymerase Kit, followed by sequencing on the PacBio Revio System using one library per SMRT Cell, in accordance with the manufacturers instructions.