GSM9339873: WT_DOX_R2; Homo sapiens; RNA-Seq
Total RNA was extracted using TRI Reagent according to the manufacturer's instructions (Merck). RNA samples were processed for rRNA removal using the Ribo-Zero kit (Illumina). Then, mRNA was randomly fragmented by adding fragmentation buffer and cDNA synthesized using random hexamers, after which a custom second-strand synthesis step was performed. This was followed by purification with AMPure XP beads, terminal repair, polyadenylation, sequencing adapter ligation, size selection and degradation of second-strand U-contained cDNA by the USER enzyme. After the final PCR enrichment, a strand-specific cDNA library was generated using the NEBNext Ultra II Directional RNA Library Prep kit for Illumina (New England Biolabs).