3.1 Transcriptome assembly and annotation
The de novo transcriptome assembly for Idiosepius pygmaeus was created from a total of 138.6 million PacBio ISO-sequencing subreads and resulted in 49,981 transcripts that were clustered into 27,420 genes. The transcriptome assembly had an N50 of 3,163 bp, 70.4% complete BUSCOs and an 82.1 ± 5.5% overall alignment rate of the RNA-seq reads (Table S3). A total of 69% of the transcripts received a functional annotation (Supplementary Table S4). The species distribution of the top blast hits was dominated by cephalopod species (Figure S22). Final mapping of RNA-seq reads against the transcriptome assembly had a 73.6 ± 6.9% mapping rate (Table S5).