2.5.1 Preparation of RNA-Seq library and Sequencing
Total RNA was extracted utilizing Trizol reagent (Invitrogen). RNA quantity and quality were determined by NanoDrop 1000 spectrophotometer (Thermo Scientific Inc.) at OD260/280, 1% agarose gel electrophoresis and Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA). Following the protocol described by Zhong et al., (2011), strand-specific RNA‐Seq libraries were prepared using 1ng/µl of total RNA sample and sequenced on Illumina HiSeq 2500 system according to the manufacturer’s instructions (Illumina).