title: GSM8398149: Acute 1; Canis lupus familiaris; RNA-Seq
library_construction_protocol: Total RNA was extracted with Trizol (Invitrogen, Carlsbad, CA, USA) the purified mRNA was used for library construction with GenSeq® Directional RNA Library Prep Kit (GenSeq, Inc., Shanghai, China) by following manufacturer's recommendations. The mRNA was fragmented into ~300 nt in length. The first-strand cDNA was synthesized from the RNA fragments by reverse transcriptase and random hexamer primers, and the second-strand cDNA was synthesized in 2nd Strand Synthesis Buffer with dUTP Mix. Then, the double stranded cDNA fragments were subjected to end-repair and dA-tailing, followed by adapter ligation. The adapter-ligated DNA samples were PCR amplified and purified to obtain sequencing libraries. Finally, the libraries were sequenced with Illumina HiSeq sequencer (Illumina, San Diego, CA, USA) on the paired-end 150 bp mode.