Deep sequencing of hepatic RNA-seq libraries from blunt snout bream
One paired-end (PE) library was generated from the pooled total RNA using an Illumina’s kit. Briefly, poly(A) mRNA was isolated from total RNA using oligo (dT) magnetic beads and then cut into short fragments in the fragmentation buffer. Taking the cleaved RNA fragments as templates, the first-strand cDNA was transcribed using random hexamer primers. The second-strand cDNA was synthesized in a mixture of buffer, dNTPs, RNaseH and DNA polymerase I. Short fragments were purified with QIAquick PCR Purification kit and eluted in QIAGEN EB buffer for end reparation and tailing A. After that, the short fragments were ligated with sequencing adapters. Following agarose gel electrophoresis, suitable fragments were selected as templates for PCR.