title: GSM6426582: Thyroid, ambient, ID 245TA; Ovis aries; RNA-Seq
library_construction_protocol: RNA was extracted using the TRIzol method (Invitrogen, Carlsbad, CA, USA) in a TissueLyser II (Qiagen, Hilden, Germany). Quality checks were done through RIN scores and the mRNA was purified from total RNA using oligo(dT)-attached magnetic beads. The resulting mRNA molecules were fragmented, and reverse transcribed into double-stranded cDNA by using random hexamer-primers. The synthesized cDNA was subjected to an end-repair and then was 3' adenylated, where adapters were ligated to the ends of these fragments. The cDNA fragments were amplified by PCR and its product was purified with Agencourt AMPure XP beads (Beckman Coulter, Brea, CA). Afterwards, a quality check was done on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The PCR product was then heat denatured and the single strand DNA was circularized by splint oligo and DNA ligase. All cDNA libraries were sequenced using paired-end strategy of 150 bp read length, through the BGISEQ-500 platform folloiwing BGI Genomics protocols.