CZ032_HIFI
PacBio HiFi sequencing is exclusively performed on the Sequel II or Sequel IIe systems (and the newer Revio system), not the outdated RS II. The library preparation involves constructing a specialized "SMRTbell" library. In this process, high-quality, high molecular weight DNA is sheared to a target size (e.g., 15-20 kb), and the fragments are end-repaired and ligated with hairpin adapters. This creates a circular, double-stranded DNA structure with single-stranded hairpin loops on both ends. During sequencing on the Sequel II system, this SMRTbell template is loaded into a Zero-Mode Waveguide (ZMW) with a polymerase. The enzyme repeatedly traverses the circular template, generating multiple, long subreads from the same molecule. These subreads are then computationally processed to produce a single, highly accurate consensus sequence known as a HiFi read, which combines the long read length of PacBio with exceptional base-level accuracy (>99.9%).