H524 SCLC cells, untreated
Messenger RNA was purified from the total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by second strand cDNA synthesis using either dUTP for the directional library or dTTP for the non-directional library. The non-directional library was prepared after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The directional library was prepared after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and a bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on an Illumina platform (NovaSeq X Plus Series PE150) according to the effective library concentration and data amount.