Isoform sequencing of Spirodela polyrhiza 7498

Single-molecule real-time (SMRT) sequencing developed by PacBio, also called third-generation sequencing (TGS), offers overwhelming longer reads than the second-generation sequencing (SGS). And isoform sequencing (Iso-Seq) of transcriptomes is advantageous for obtaining full-length transcripts without assembly and could be used for genome annotation, identification of novel genes and isoforms, as well as the discovery of long non-coding RNA (lncRNA). Here, we we sequenced and characterized the full-length transcriptomes of Spirodela polyrhiza 7498 using the PacBio long-read sequencing technology. Four libraries including 1-2 kb, 2-3 kb, 3-6 kb, and 5-10 kb were constructed using BluePippin size selection system. The libraries were sequenced on the PacBio Sequel system with 4 SMRT cells, yielding 1,606,022 polymerase reads with the average length of over 10 kb. There were 545,753 full-length non-chimeric (FLNC) reads which contained 5' primer, 3' primer and the poly (A) tail. The length distributions of ROIs were expected as each library established, with an average of 1,292, 2,225, 3,344 and 5,715 bp, respectively. The full-length non-chimeric reads were further correction and and a total of 492,435 high quality FLNC (HQFLNC) were obtained.