Not applicable
Briefly, polyadenylated mRNA was isolated from 1g of total RNA using oligo(dT) magnetic beads. Purified mRNA was fragmented at 94C for 8 minutes and subsequently reverse-transcribed into first-strand cDNA using random hexamer primers and SuperScript II Reverse Transcriptase (Invitrogen, Carlsbad, CA, USA). Second-strand cDNA synthesis was performed to generate double-stranded cDNA. Final libraries were quantified by qPCR and analyzed for size distribution using the Agilent Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) prior to sequencing. A total of 6 libraries were sequenced using a paired-end strategy (2 x 150 bp) with the NovaSeq6000 Illumina platform of Macrogen (Seul, Korea).