RNA-seq of equus caballus: adult male  skelental muscle
Total RNA from twelve samples (three  replicates from each of four parts ) were extracted using samples in -80  above following the manufacturer's procedure of Trizol reagent (Invitrogen,  CA, USA) and Animal Tissue RNA Purification Kit, TRK1002 (LC Science,  Houston, TX). The total RNA quantity and purity were analysis of Bioanalyzer  2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA) with RIN number  >7.0. Approximately 10 ug of total RNA representing a specific adipose  type was used to deplete ribosomal RNA according to the manuscript of the  Epicentre Ribo-Zero Gold Kit (Illumina, San Diego, USA). Following  purification, the poly(A)- or poly(A)+ RNA fractions is fragmented into small  pieces using divalent cations under elevated temperature. Then the cleaved  RNA fragments were reverse-transcribed to create the final cDNA library in  accordance with the protocol for the mRNA-Seq sample preparation kit  (Illumina, San Diego, USA), the average insert size for the paired-end  libraries was 300 bp (50 bp). And then we performed the paired-end sequencing  on an Illumina Hiseq 4000.