GSM9745198: NESCs, HT cell line, aged, biological replicate 3; Homo sapiens; RNA-Seq
Total RNA was extracted from NESCs and snap-frozen. RNA quantity and integrity were assessed using the Qubit RNA High Sensitivity Assay Kit (Invitrogen) and the High Sensitivity RNA ScreenTape system (Agilent Technologies). Polyadenylated mRNA was isolated using paramagnetic bead-based capture. Library preparation was performed using the AccuraCode RNA-Seq Kit V2 (Singleron Biotechnologies GmbH) according to the manufacturer's instructions. During reverse transcription, 100 ng mRNA was converted to cDNA and labeled with sample-specific barcodes and unique molecular identifiers. Barcoded cDNA from all samples was pooled and amplified. A total of 50 ng cDNA was used for library construction. Libraries were sequenced on a NovaSeq X platform (Illumina) using paired-end 150-bp sequencing by GENEWIZ (Leipzig, Germany).