RNA-seq of human iPSC-derived astrocytes, control brain treated
Total RNA was extracted from iPSC-derived astrocytes using a standard TRIzol-based protocol. Poly-A RNA selection was performed to isolate mRNA from the total RNA using oligo(dT) beads, ensuring enrichment of coding RNA and removal of ribosomal RNA. The enriched mRNA was then fragmented and reverse-transcribed to synthesize cDNA. The cDNA underwent end repair, A-tailing, and adapter ligation for library preparation. PCR amplification was performed to generate the final sequencing library. Quality control of the RNA and library was assessed using an Agilent Bioanalyzer, and the concentration was quantified by Qubit. The libraries were sequenced using the Illumina NovaSeq 6000 platform with a 2x150 bp paired-end configuration, providing comprehensive coverage of the transcriptome