PacBio_Hifi_reads_M_telemachus_gold
Library preparation and sequencing were performed according to the manufacturers instructions. At each step, DNA was quantified using the Qubit dsDNA HS Assay Kit. DNA purity was tested using the nanodrop and size distribution and degradation assessed using the Femto pulse Genomic DNA 165-kb Kit. Purification steps were performed using AMPure PB beads. Next, 15g DNA was purified and then sheared at 15 kb with the Megaruptor3 system. Using the SMRTbell Express Template prep kit 2.0, a single-strand overhang removal, a DNA and END damage repair step was performed on 10g of sample. Blunt hairpin adapters were then ligated to the library, which was treated with an exonuclease cocktail to digest unligated DNA fragments. A size selection step using a 10-kb cutoff was performed on the BluePippin Size Selection system. Using Binding kit 2.2 and sequencing kit 2.0, the primer V5 annealed and polymerase 2.2 bounded library was sequenced by diffusion loading onto 1 SMRTcells per sample.