Charactering the transcriptome of Abramis brama (Cyprinidae: Abramis) enables ecological and evolutionary analyses on the species
A total of 0.75µg total RNAs from each sample were mixed and used for subsequent library construction and sequencing. First of all, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Then, fragmentation of mRNA was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). After that, random hexamer primer was used to reverse-transcribe the mRNA fragments and cDNA fragments of preferentially 150~200bp in length were selected. After PCR amplification and purification, quality of the cDNA library was assessed on an Agilent Bioanalyzer 2100 system.