title: RNA-seq of equus caballus: adult male skin
design: 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, TRK1014 (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.