GSM8833564: NR-001-iA_C; Homo sapiens; RNA-Seq
Total RNA was extracted from each of the nine hiPSC-derived astrocyte cell lines via TRIzol reagent (Invitrogen, 15596026) per the manufacturer's instructions. Total RNA from each line was treated with DNase I (Zymo Research, E1010) and then cleaned and concentrated via the Clean & Concentrator Kit (Zymo Research, D4033). A NanoDrop One/OneC Microvolume UV‒Vis Spectrophotometer (Thermo Scientific, ND-ONE-W) was then used to determine the RNA yield from each of the nine RNA extractions. For RNA sequencing (RNA-seq), 150-cycle PE sequencing via the Illumina HiSeq2500 platform was performed by Admera Health (South Plainfield, New Jersey, United States) with 1 µg of RNase-free RNA (>10 ng/μL, RIN > 7) per technical triplicate for each of the nine lines. An RNA-seq library was generated utilizing the Illumina mRNA sample prep kit (Illumina, RS-100-0801) according to the manufacturer's protocol in 3 technical replicates for each sample. The library constructs' size and concentration were verified with a bioanalyzer before sequencing on the Illumina NovaSeq6000. The raw reads' quality was evaluated to confirm the success of the library preparation and sequencing processes.