Full length transcriptome sequencing of mud crab crablet
Briefly, the SMARTer PCR cDNA Synthesis Kit with Oligo-dT primers was used to generate first- and second-strand cDNA from polyA mRNA. Size fractionation and selection were performed using the BluePippin Size Selection System. Two SMRT bell libraries were constructed with the Pacific Biosciences DNA Template Prep Kit 2.0 and SMRT sequencing was then performed using the Pacific Bioscience Sequel System. Full-length sequencing and analysis pipeline. We combined all raw data and performed initial data processing according to the Iso-seq standard pipeline. The Circular consensus sequence (CCS) was generated from initial data using the SMRTlink (version 5.1) software16. The CCS was classified into full-length and non-full length reads according to the 5and 3adapters and the poly(A) tail. Reads containing both the 5 and 3 primers and having a poly(A) tail signal preceding the 3 primer were considered to be full-length reads.