Total DNA quality was analysed on a PacBio RS II sequencing platform analyser system and by resolution on a denaturing polyacrylamide gel electrophoresis system. A DNA database library was generated according to the DNA sample preparation instructions. Subsequently, DNA was amplified with Pfx DNA polymerase (Invitrogen, China) using 20 PCR cycles and a PacBio DNA primer set. PCR products were purified, and the recovered DNA was precipitated and quantified with both a Nanodrop Spectrophotometer (Thermo Scientific) and a TBS-380 mini fluorometer (Turner Biosystems) using PicoGreenH dsDNA quantitation reagent (Invitrogen). The sample concentration was adjusted to 10 nM, and a final volume of 10 mL was used for the sequencing reaction. The purified DNA library was used for cluster generation (on the PacBio Cluster Station). Subsequently, DNA was sequenced on a PacBio Sequel machine following the manufacturer's instructions (Nextomics), and the library construction process is shown in Fig. 1 . The gDNA concentration was normalized by dilution from a high to a low concentration and was then sequenced using the PacBio platform.Fig. 1 ![]()
The genomic DNA library was generated according to the PacBio Sequel sample preparation instructions.