GSM8075825: iPSC-derived motor neuron cultures, control, line EDi022-A, Chip, Static, rep 2, experiment 1.3; Homo sapiens; RNA-Seq
Total RNA samples were assessed for concentration using a Qubit fluorometer (ThermoFisher Scientific, Waltham, MA) and for quality using the 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).  Up to one µg of total RNA was purified for mRNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolab Inc, Ipswich, MA).  Stranded RNA-Seq library construction was performed using the IDT xGen Broad-Range RNA Library Prep Kit (Integrated DNA Technologies, Coralville, IA).  Library concentration was measured with a Qubit fluorometer and library size on an Agilent 4200 TapeStation (Agilent Technologies). Libraries were multiplexed and sequenced on a NovaSeq 6000 (Illumina, San Diego, CA) using 75bp single-end sequencing. On average, approximately 30 million reads were generated from each sample. The top channel of chip samples or a well from a 96 well plate were incubated with cold Accutase (STEMCELL Technologies) for 20 minutes at 37°C, then cells were extracted by flushing and washing with cold phosphate buffered saline (PBS), pelleted by centrifuge at 350 rcf, and frozen at -80°C. RNA was extracted from pellets using the RNeasy Micro Kit (Qiagen). Various methods:  see SAMPLES "library construction protocol" section