Transcriptome sequencing, de novo assembly and genes annotation of samples from different tissues of Chionodraco hamatusi
The extraction of total RNA was carried out using Trizol (Invitrogen, CA, USA) according to the manufacturer’s instruction. The quantity and purity of the extracted RNA were analyzed using Bioanalyzer 2100 and RNA 6000 NanoLabChip Kit (Agilent, CA, USA) with RIN number >8.0. Approximately 10 ug of total RNA representing a distinctive adipose type was employed for Poly (A) mRNA isolation using poly-T oligo attached magnetic beads (Invitrogen). Subsequent to purification, the mRNA wasbroken into small pieces using divalent cations at high temperature. The sliced RNA fragments were reverse-transcribed forgenerating the cDNA library according to the mRNA-Seq sample preparation kit (Illumina, San Diego, USA) instructions. The paired-end sequencing (2*150 bp) on an Illumina Hiseq4000 was implemented according to the manufacturer procedure using paired-end libraries with normal insert size of 300 bp (±50 bp).