Iso-Seq for gene annotation.
Total RNA was isolated  according to the instruction of Trizol method. The integrity, quality and concentration of RNA were detected by NanoDrop 2000 spectrophotometer (Thermo Fischer Scientific) and Angilent 2100 Bioanalyzer (Agilent Technologies). High quality RNA (2g, 300ng/l) was prepared for the next step of library construction.