GSM7698110: WT_CTRL_MN1; Homo sapiens; RNA-Seq
RNA from Day 10 NPCs, Day 28 motor neurons and D35 astrocytes was extracted using the RNeasy Mini Kit (Qiagen, Germany, #74104) with RNA clean-up. Day 28 motor neurons were enriched for neurons by magnetic cell sorting using MACS cell separation antibodies, microbeads, and reagents (Miltenyi Biotec) targeting CD171+-PSA+-NCAM+ cells. The mRNA is fragmented randomly by adding fragmentation buffer, then the cDNA is synthesized by using mRNA template and random hexamers primer, after which a custom second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I are added to initiate the second-strand synthesis. Second, after a series of terminal repair, a ligation and sequencing adaptor ligation, the double-stranded cDNA library is completed through size selection and PCR enrichment.