title: RNA-Seq  of Sillago sihama: normoxia-treated fish gill bioreplicate 1
design: Fragmentation was carried out using divalent  cations under elevated temperature in NEBNext First Strand Synthesis Reaction  Buffer(5X). First strand cDNA was synthesized using random hexamer primer and  M-MuLV Leading Edge Genomic Services & Solutions Reverse  Transcriptase(RNase H - ). Second strand cDNA synthesis was subsequently  performed using DNA Polymerase I and RNase H. Remaining overhangs were  converted into blunt ends via exonuclease/polymerase activities. After  adenylation of 3 ends of DNA fragments, NEBNext Adaptor with hairpin loop  structure were ligated to prepare for hybridization. In order to select cDNA  fragments of preferentially 250~300 bp in length, the library fragments were  purified with AMPure XP system