sample treatment protocol
nucleic acid library construction protocol
RNA was extracted using NucleoSpin RNA kit, Macherey-Nagel. All samples exhibited a RIN value of >7. Library preparation for RNA-Seq was performed using the TruSeq Stranded Total RNA Sample Prep Kit, Illumina starting from 500 ng of total RNA according to Illumina's instructions. Accurate quantitation of cDNA libraries was performed by using the QuantiFluor™ dsDNA System (Promega). The size range of final cDNA libraries was determined applying the DNA 1000 chip on the Bioanalyzer 2100 from Agilent (280 bp). The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the HiSeq2000 following the manufacturer's protocols.