Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: NSCs were collected after 4 days and astrocytes were collected after 6 days of culture. Cells were lysed in RLT buffer supplemented with 2-mercaptoethanol. Cell lysates were stored at -80C before RNA extraction.Total RNA was extracted with RNeasy Mini kit (Qiagen) following manufacturer protocol. RNA (200 ng) was used to prepare cDNA libraries using the TruSeq Stranded mRNA HT Library Preparation Kit (Illumina). Libraries were normalized, pooled and then clustered using the HiSeq® 3000/4000 PE Cluster Kit (Illumina). The libraries were imaged and sequenced on an Illumina HiSeq 4000 sequencer using the HiSeq® 3000/4000 SBS kit (Illumina) at a minimum of 25 million single-end reads (76 bp) per sample.