PACBIO reads Phillyrea angustifolia 01N25
High Fidelity (HiFi) libraries were constructed using the SMRTbell Template Prep kit 2.0 (Pacific Biosciences, Menlo Park, CA, USA) according to PacBio recommendations (SMRTbell express template prep kit 2.0 - PN: 100-938-900). HMW DNA samples were first purified with 1X Agencourt AMPure XP beads (Beckman Coulter, Inc, CA USA), and sheared with Megaruptor 3 (Diagenode, Lige, BELGIUM) at an average size of 20 kb. After end repair, A-tailing and ligation of SMRTbell adapter, the library was size-selected on the BluePippin System (Sage Science, MA,USA) at range sizes of 10-50kb. The size and concentration of libraries were assessed using the Agilent FP-1002 Genomic DNA 165 kb on the Femto Pulse system and the Qubit dsDNA HS reagents Assay kit.Sequencing primer v5 and Sequel II DNA Polymerase 2.2 were annealed and bound, respectively, to the SMRTbell libraries. Each library was loaded on two SMRTcell 8M at an on-plate concentration of 90pM. Sequencing was performed on the Sequel II system at the Gentyane Genomic Platform (INRAE Clermont-Ferrand, France) with Sequel II Sequencing kit 3.0, a run movie time of 30 hours with an Adaptive Loading target (P1 + P2) at 0.75.