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).