PacBio HiFi of M. javanica VW4
HiFi SMRTbell libraries were constructed using the SMRTbell Express Template Prep Kit v2.0 (Pacific Biosciences, Menlo Park, CA; Cat. #100-938-900) according to the manufacturer's instructions. HMW gDNA was sheared to a target DNA size distribution between 15 kb  20 kb using Diagenodes Megaruptor 3 system (Diagenode, Belgium; Cat. B06010003). The sheared gDNA was concentrated using 0.45X of AMPure PB beads (Pacific Biosciences, Menlo Park, CA; Cat. #100-265-900) for the removal of single-strand overhangs at 37 C for 15 minutes, followed by further enzymatic steps of DNA damage repair at 37 C for 30 minutes, end repair and A-tailing at 20 C for 10 minutes and 65 C for 30 minutes, ligation of barcoded overhang adapters v3 at 20 C for 60 minutes and 65 C for 10 minutes to inactivate the ligase, then nuclease treated at 37 C for 1 hour. SMRTbell libraries were purified and concentrated with 0.45X Ampure PB beads for size selection using the BluePippin/PippinHT system (Sage Science, Beverly, MA; Cat #BLF7510/HPE7510) to collect fragments greater than 7-9 kb. HiFi SMRTbell libraries were sequenced at UC Davis DNA Technologies Core (Davis, CA) using one SMRT Cell 8M Tray (Pacific Biosciences, Menlo Park, CA; Cat #101-389-001), Sequel II sequencing chemistry 2.0, and 30-hour movies each on a PacBio Sequel II sequencer.