design: The PacBio HiFi library preparation, known as the SMRTbell workflow, begins with the shearing of high-molecular-weight genomic DNA into specific target sizes (typically 1520 kb), followed by enzymatic end-repair and dA-tailing. Specialized hairpin adapters are then ligated to both ends of the double-stranded DNA fragments, creating a structurally linear but topologically circular DNA template called a SMRTbell. After purification and size selection using magnetic beads or pulsed-field electrophoresis to remove short fragments, a sequencing primer and DNA polymerase are bound to the adapter regions. During sequencing, the polymerase continuously circles the template, generating multiple passes of the same molecule (subreads), which are then computationally combined to produce a Highly Fidelity (HiFi) consensus read with accuracy exceeding 99.9%.