PacBio HiFi whole genome sequencing of Neodiprion virginiana: eonymph male thorax and abdomen
High molecular weight (HMW) DNA extraction was performed from the thorax and abdomen using the Qiagen MagAttract HMW DNA Kit. DNA shearing was performed using the Diagenode Megaruptor 2 with the 20 kb fragment protocol. The sheared DNA was prepared for PacBio sequencing using the SMRTBell Express Template kit 2.0. The library was size-selected with beads to remove library molecules less than 3 kb in length to generate the final libraries for sequencing. Sequencing was performed on a Sequel II System using Binding Kit v2.0, Sequencing kit v2.0, and SMRT Cell 8M. To target HiFi reads, the library was sequenced using a 30 hour movie time on one SMRTcell. Raw subreads were converted to HiFi data by processing with CCS to call a single high quality consensus sequence for each molecule, using a 99.5% consensus accuracy cutoff.