2 cm female cones
Total RNA was isolated by scaling down and adapting the method CTAB-based lysis solution with the silica column-based RNA binding, DNase, and washing steps from an RNeasy Plant Mini Kit. Iso-Seq libraries were constructed following the PacBio modified protocol using the Clonetech SMARTer PCR cDNA Synthesis Kit and Blue Pippin Size Selection System. Libraries were then prepared using SMRTbell library protocol. Each library was sequenced across 4 SMRT cells on the Pacific Biosciences RSII using P6-C4 chemistry. 3-6 Kb size selected library.