PacBio HiFi sequencing of Clemmys guttata: adult female blood
A HiFi SMRTbell library was constructed using a SMRTbell prep kit (3.0; PacBio) following the manufacturers protocol. DNA was sheared to a target size distribution between 15-18 kb using a Megaruptor 3 system (Diagenode, Belgium). Sheared DNA was concentrated using 1X SMRTbell cleanup beads provided in the SMRTbell prep kit for the repair and a-tailing incubations at 37C for 30 minutes and 65C for 5 minutes, respectively, followed by ligation of overhang adapters at 20C for 30 minutes, clean-up using 1X SMRTbell cleanup beads, and nuclease treatment at 37C for 15 minutes. The SMRTbell library was size selected using 3.1X of 35% v/v diluted AMPure PB beads (PacBio) to progressively remove SMRTbell templates <5 kb. The 15-18 kb average HiFi SMRTbell library was sequenced using one 25M SMRT cell (PacBio), Revio sequencing chemistry 2.0, and 30-hour movies on a PacBio Revio sequencer.
SUB15684660
CLGU-HIFI1