title: Pacbio HiFI seq of the Cibotium barometz: young fronds
design: DNA extraction and HiFi library preparation High-molecular-weight DNA was extracted from young fronds of Cibotium barometz using a modified CTAB method. Fresh tissue was ground in liquid nitrogen, and DNA was isolated using CTAB buffer, followed by chloroform extraction and isopropanol precipitation. DNA quality and quantity were assessed using Nanodrop spectrophotometry and pulsed-field gel electrophoresis.HiFi sequencing libraries were prepared following the SMRTbell prep kit 3.0 protocol (Pacific Biosciences). Briefly, DNA was sheared to ~15-20 kb fragments using a Megaruptor 3 system (Diagenode). The sheared DNA underwent damage repair and end-repair/A-tailing steps, followed by adapter ligation. Size selection was performed using a BluePippin system (Sage Science) to enrich for 15-20 kb fragments. The final library was assessed for quality and quantity using a Femto Pulse system (Agilent).Sequencing HiFi sequencing was performed on a PacBio Sequel IIe platform using one SMRT cell. Sequencing was conducted with 30-hour movie times to generate HiFi reads with 15-20 kb insert sizes and Q20 or greater base quality. Data processing Raw sequencing data were processed using the SMRT Link software (v10.1) to generate high-quality HiFi reads.