PacBio HiFi Sequencing of Nannochloropsis oceanica QH25
The gDNA was sheared to the target size of 15 kbp on Megaruptor 2 and purified with AMPure PB beads. HiFi SMRT libraries were prepared according to the Pacific Biosciences protocol. Single strand overhangs were removed, followed by the DNA Damage repair, End repair and A-tailing reactions. Overhang adapters were ligated, followed by a nuclease treatment reaction to remove damaged DNA and misligated products. For sequencing on the Sequel instrument the SMRT bell libraries were purified and size selected using diluted AMPure PB beads. Sequencing primer v.4 was annealed and Sequel DNA polymerase 3.0 bound to the SMRT bells. Diluted SMRT bell/ DNA polymerase complex was sequenced on two SMRT cells 1M using the sequencing plate 3.0 and 20 hour movies.