title: GSM8242184: BTS11_C1_RNA-seq; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was extracted pellets of 100,000 cells in 400 μL of TRIzol (Invitrogen) and phase-separated by adding 80 μL of chloroform:isoamyl alcohol (Sigma-Aldrich) after centrifugation at 4 °C for 15 minutes. The aqueous phase was mixed with 1 μL of GlycoBlue and 300 μL of cold isopropanol and incubated on ice for 10 minutes. RNA was pelleted by centrifugation for 10 minutes at 4 °C, washed with 75% ethanol, air-dried, and resuspended in 5 μL of RNase-free water. Oligo (dT)25 capture of mRNA was done using Dynabeads mRNA DIRECT kit (Life Technologies), with previously described adaptations (Hanna et al., 2019). The SMARTer Stranded RNA-seq kit (Takara) was used to generate libraries from isolated mRNA, using 12 amplification cycles. Libraries were then quantified using the High DNA Sensitivity Bioanalyzer 2500 (Agilent) and Illumina library quantification kit (KAPA).