GSM8645920: 0.4 µm-pored membrane, biol rep 1; Homo sapiens; RNA-Seq
RNA was extracted using the RNeasy Mini kit (Qiagen, Hilden, Germany). mRNA was enriched using oligo(dT) beads. The mRNA was fragmented randomly by adding a fragmentation buffer, then the cDNA was synthesized using an mRNA template and a random hexamer primer, after which a custom second-strand synthesis buffer (Illumina), dNTPs, Rnase H, and DNA polymerase I were added to initiate second-strand synthesis. After a series of terminal repair, A-tailing, and sequencing adaptor ligation, the double-stranded cDNA library was completed through size selection and PCR enrichment.