Isoform Sequencing of M. hapla
cDNA and SMRTbell library were constructed using SMRTbell prep kit 3.0 (PN 102-396-000 REV02 APR2022) according to manufacturers instructions. The cDNA was synthesized using NEBNext Single Cell/Low Input cDNA Synthesis & Amplification Module (New England Biolabs, Ipswich, MA; Cat. #E6421L). After fifteen cycles of PCR for cDNA amplification, the cDNA was purified using 0.86X SMRTbell cleanup beads. Subsequently, the cDNA libraries derived from each sample were pooled, and used for SMRTbell library construction using the SMRTbell prep kit 3.0 (Pacific Biosciences, Menlo Park, CA; Cat. #102-182-700) with the following enzymatic steps: repair and a-tailing, ligation of overhang adapters, and nuclease treatment. The Iso-Seq SMRTbell library was sequenced at UC Davis DNA Technologies Core (Davis, CA) using one 8M SMRT cell (Pacific Biosciences, Menlo Park, CA; Cat #101-389-001), Sequel II sequencing chemistry 2.0, and 24-hour movies on a PacBio Sequel II sequencer.