Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Cell pellets were resuspended in 1 ml of TRIzol and stored at -80˚ C. The sample was then heated for 10 min at 65°C and 200 µl of chloroform was added, mixed and incubated at room temperature for 5 min. Samples were centrifuged at 15,000 × g for 15 min at 4°C. The clear aqueous phase was transferred to a new tube and 0.7 volume of cold 100% isopropanol was added. Samples were then saved at -80°C overnight. The overnight precipitation was centrifuged at 15,000 × for 30 min at 4°C and the resulting nucleic acid pellet was washed twice with cold 70% ethanol. The pellet was centrifuged again at 15,000 × for 30 min at 4°C, ethanol was removed and the pellet was allowed to dry. The nucleic acid pellet was resuspended with nuclease-free water. The samples were treated with with Turbo DNase (Invitrogen) and clean-up was performed with RNeasy Mini Kit (Qiagen). TruSeq stranded total RNA kit, with ribo-zero ribosomal RNA reduction was used to prepare RNA-seq library, per manufacturer (Illumina) protocol