WT-1
Poly(A)+ RNA (messenger RNA [mRNA]) was enriched using magnetic beads with oligo (DT) and fragmented using a fragmentation buffer. After single-stranded complementary DNA (cDNA) synthesis with random hexamers, buffer, dNTPs, DNA polymerase I, and RNase H were used to synthesize double-stranded (ds) cDNA. The acquired ds cDNA was purified with AMPure XP beads (Beckman Coulter, Brea, CA, USA), followed by end-repaired, 3 ends adenylated, and barcoded adaptor ligated to the end. Fragments were enriched by PCR to obtain the final libraries, followed by primary quantification using Qubit 2.0 dsDNA High Sensitivity Assay Kit (Thermo Fisher Scientific). The diluted libraries were accurately assayed using qRT-PCR and sized using an Agilent 2100 Bioanalyzer (Agilent Technologies).