WGS_HiFi
Genomic DNA (gDNA) was sheared using a g-TUBE to fragments between 6 and 20 kb for 10 kb and 20 kb SMRTbell library construction. DNA with long overhangs was treated with ExoVII before damage repair. Repair enzymes from the Template Prep Kit were used to fix various DNA damages. T4 DNA Polymerase filled 5 overhangs and removed 3 overhangs, while T4 PNK phosphorylated 5 ends. Hairpin adapters were ligated to the repaired ends. Excess and imperfect SMRTbells were removed with EXOIII and ExoVII. AMPure PB Beads purified the library, which was further size-selected using the BluePippin System for large fragments. The selected SMRTbell templates were then purified again with AMPure PB Beads. A sequencing primer was attached, and a polymerase bound to the templates for loading into Zero-Mode Waveguides (ZWMs) using the Binding Kit. Finally, libraries were loaded into SMRT Cells, and the SMRT Cell 8M Tray was used for sequencing.