RNA-ref of Megalobrama amblycephala: skin
Oligo(dT)-attached magnetic beads were used to purify the mRNA. Purified mRNA was fragmented into small pieces with fragmenting buffer at appropriate temperature. First- and second-strand cDNA was generated by PCR. The reaction products were purified using magnetic beads afterwards. A-Tailing Mix and RNA Index Adapters were added by incubating to end repair. The cDNA fragments with adapters were amplified by PCR, and the products were purified by Ampure XP Beads. Quality and quantity of the library were assessed using the Agilent 2100 bioanalyzer. The qualified library was amplified on cBot to generate the cluster on the flowcell. The amplified flowcell was sequenced (single end) on the HiSeq4000 or HiSeq X-ten platforms