PacBio sequencing of Hirschfeldia incana: leaf
SMRTbell preparation was performed based on the PacBio SMRTbell Express Template Kit v2.0 protocol using 3 g DNA input. The final library was size selected using the Blue Pippin protocol for fragments >13 kb. Primer (Sequencing Primer v4) and polymerase were bound to the SMRTbell using the Sequel Binding and Internal Ctrl Kit 3.0 to form a SMRTbell complex. Purification of the SMRTbell complex was performed following the Pacbio Procedure & Checklist  AMPure PB Bead Purification of Polymerase Bound SMRTbell Complexes to remove excess polymerase and primers. Prior to sequencing a DNA Internal Control Complex v3 was added at a low percentage to the diluted complex according to protocol. Finally, sequencing was performed using diffusion loading on the Sequel SMRT Cell 1M v3 and Sequel Sequencing Kit 3 reagents.