title: GSM5420357: 010.85-Mobs; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was isolated using the RNeasy kit from Qiagen according to the manufacturer's protocol. Concentrations of RNA samples were quantified using the NanoDrop 2000 spectrophotometer (Thermofisher) according to the manufacturer's protocol. Quality was assessed by computing RNA integrity numbers (RINs) using the Agilent 2100 Bioanalyzer according to the manufacturer's instructions. Total RNA (800 ng) was ribosomal RNA (rRNA) depleted using the Low Input RiboMinus™ Eukaryote System v2 (Thermofisher) according to the manufacturer's instructions. Samples were then vacuum concentrated and whole-transcriptome and barcoded libraries were prepared using the Ion Total RNA-Seq Kit v2 (Thermofisher) according to the manufacturer's instructions. Size distribution of samples was assessed using the Agilent 2100 Bioanalyzer and 6000 RNA Pico kit. Library concentrations were quantified using the Agilent DNA 1000 assay and the 2100 Bioanalyzer according to the manufacturer's instructions. Barcoded whole-transcriptome libraries were diluted to a 66 pM concentration and combined in equal volumes (13 µl each, 26 µl total) for sequencing with two samples per template preparation. All template reactions were performed on the Ion Chef Instrument using the Ion PI Hi-Q Chef kit (Thermofisher) according to the manufacturer's instructions. Reactions were loaded onto Ion PI Chips v3 (Thermofisher) for sequencing on an Ion Proton sequencer according to the manufacturer's protocol.