PacBio HiFi
The Pacific Biosciences protocol Preparing whole genome and metagenome libraries using SMRTbell prep kit 3.0 was used to create a library between 5-7.5 micrograms of DNA. The Megaruptor (Diagenode) was used for shearing and Blue Pippin (Sage Science) was used for size-selection for fragments ~11-17kb. Library size was assessed using a FemtoPulse (Agilent). Each library was run on 25M SMRTCells using SPRQ sequencing reagents. Sequencing was performed on a Revio sequencer (Pacific Biosciences) running instrument control version 13.3.0.253824 and a movie collection time of 30 hours per SMRTCell with adaptive loading. Demultiplexed CCS/HiFi unaligned bam files with kinetic information and 5mC tags (called with jasmine) were generated on-instrument using tools in software version 13.3.0.253824.