Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNA was isolated and purified using the QIAGEN RNeasy Mini Kit. Lysozyme and lysostaphin were used for cell wall degradation, followed by two cycles of two-minute bead beating with 1 ml of 0.1 mm silica beads in a mini bead-beater (BioSpec), and eluted in nuclease-free water. Isolated RNA was treated with 1 uL of Baseline Zero DNase (Epicentre) at 37°C for 30 min and ribosomal RNA depletion was performed using Epicenter Ribo-ZeroTM Magnetic Gold Kit (Illumina), according to the manufacturer’s instructions. After RNA extraction, barcoded stranded RNA-Sequencing libraries were prepared using the TruSeq Stranded Total RNA Sample Preparation kit (Illumina). RNA quality and quantity was assessed using the Agilent Bioanalyzer and Qubit RNA Broad Range Assay kit (Thermo Fisher), respectively. Finally, Libraries were pooled and sequenced on the Illumina HiSeq platform in a 100 bp single-end read run format.