PacBio Sequencing for Phytophthora nicotianae race 1
Genomic DNA samples were first sheared to target insert size (10 kb) using the Covaris® g-TUBE. Fragmented DNA was then purified using AMPure® PB magnetic beads. Libraries were subsequently prepared following PacBio guidelines. SMRTBell libraries were verified using Life Technologies Qubit® 2.0 and the Agilent Bioanalyzer. The PacBio specific sequencing primer was annealed to the SMRTbell library followed by binding of the polymerase to the primer library complex. Libraries were loaded onto the SMRT cells with the assistance of the MagBead stations and sequenced on the PacBio RS II system. Libraries were sequenced with stage-start settings, and used 1×120 movies. Both libraries were sequenced using XL binding kit and C2 sequencing kit.