title: GSM8682646: organoid microglia, C9-1, biol rep1; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated using QIAzol Lysis reagent (Qiagen) and miRNeasy mini kit (Qiagen) with DNA digestion on-column, according to the manufacturer's instructions. RNA concentration and integrity (RIN) was verified in an Agilent 2100 bioanalyzer and only samples with RIN ≥ 9.0 were used for RNA sequencing. Sample preparation was performed according to "NEBNext Ultra II Directional RNA Library Prep Kit for Illumina" (NEB #E7760S/L). Briefly, mRNA was isolated from total RNA using the oligo-dT magnetic beads. After fragmentation of the mRNA, a cDNA synthesis was performed. This was used for ligation with the sequencing adapters and PCR amplification of the resulting product. The quality and yield after sample preparation was measured with the Fragment Analyzer. The size of the resulting products was consistent with the expected size distribution (a broad peak between 300-500 bp). Samples were sequenced using the Illumina NovaSeq 6000, following a data quality metrics check for possible sample and barcode contamination, using third-party (FastQC v0.11.9) and in-house (FastQA v3.1.25) quality control tools.