Full-length transcriptome sequencing of the veined rapa whelk
Larval development stages were sampled: one–spiral whorl stage, 1 day post-hatching (dph), two–spiral whorl stage (5 dph), three–spiral whorl stage (18 dph), and four–spiral whorl stage (competent larvae, 35 dph). Samples were also collected from the post-larval stage (33 dph, 3 days after settlement) and adults. The samples were washed with ddH2O and snap frozen in liquid nitrogen until use. Samples from all six developmental stages were used for RNA extraction using a Trizol Kit (Invitrogen, USA) following the manufacturer's instructions. The purity and integrity of the RNAs were checked using a NanoPhotometer® spectrometer (IMPLEN, CA, USA) and 1.1% agarose gel electrophoresis The RNA concentration was measured using a Qubit® RNA Assay Kit in a Qubit® 2.0 Fluorometer (Life Technologies, CA, USA). Equal amounts of RNA from each stage were pooled into a single library for synthesizing double-stranded cDNAs. A PacBio Template Prep Kit (PacBio, CA, USA) was then used to generate three SMRT bell libraries. The SMRT bell templates were sequenced on the PacBio Sequel System