GSM8075874: iPSC-derived motor neuron cultures, sporadic ALS, line 5XVDiALS, Chip, Flow, rep 2, experiment 5.4; Homo sapiens; RNA-Seq
RNA integrity was analyzed on the 2100 Bioanalyzer using the Agilent RNA 6000 Nano Kit (Agilent Technologies, Santa Clara, CA) and RNA quantified using the Qubit RNA HS Assay Kit (ThermoFisher Scientific, Waltham, MA).  Total RNA-Seq library construction was performed using the IDT xGen Broad-Range RNA Library Prep Kit (Integrated DNA Technologies, Coralville, IA), following rRNA depletion using Lexogen RiboCop Depletion Kit Human/Mouse/Rat v2 (Lexogen Inc., Greenland, NH).  Final library concentration was measured via the Qubit 1X dsDNA HS Assay kit (ThermoFisher Scientific) and library size was evaluated on the 4200 TapeStation using the Agilent D1000 ScreenTape System (Agilent Technologies).  Multiplexed libraries were sequenced on a NovaSeq 6000 (Illumina, San Diego, CA) using 75bp single-end sequencing.  On average, about 50 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