GSM7732703: RNA-seq_ER232-1_ITD-16; Homo sapiens; RNA-Seq
RNA extraction was done using a RNeasy Micro Plus kit (QIAGEN) where less than 50,000 cells were harvested, and a RNeasy Micro Plus kit (QIAGEN) for larger cell numbers. RNA was quantified by nanodrop and QC'd using an Agilent RNA 6000 Pico Kit (Agilent, bioanalyser). Libraries for next generation sequencing were prepared using the NEBnext Ultra II Directional RNA Library Prep Kit for Illumina (NEB) with the NEBNext® rRNA Depletion Kit v2 for low RNA input (<100 ng RNA), or the Total RNA Ribo-zero library preparation kit (with ribosomal RNA depletion) (Illumina) for higher RNA input. Libraries were quantified using the High Sensitivity DNA kit (Agilent) and Kapa Library Quantification kit (Roche) prior to paired end sequencing on a Next Seq 2000 (PE 75) with a NextSeq High 150 v2.5 kit.