GSM8921754: HK-244 cells 0 Gy Control; Homo sapiens; RNA-Seq
At day 5, the gliomaspheres in the suspension culture were collected and dissociated with TrypLE (no phenol red), and the adherent differentiated cells were washed with HBSS and de-attached with Trypsin/EDTA. The cells were then pooled and filtered through a 40-µm strainer and counted with a hemacytometer. The samples were pelleted by centrifugation at 300 g for 5 minutes at 4°C, resuspended in the provided Cell Resuspension Buffer (Fluent Bioscience, Watertown, MA), and 50,000 cells were targeted for capture and processed for single-cell transcriptomics using the PIPseqTM T2 3' Single Cell RNA Kit V5 (Fluent Biosciences). Sequencing libraries were prepared according to PIPseq T2 instructions, with 12 cycles for cDNA amplification and 8 cycles for library amplification, depending on the DNA amount. The libraries were quantified using a Qubit 4 Fluorometer and a TapeStation 4200 and sequenced as per manufacturer recommendation on a NovaSeq X Plus – 25B 2 x150bp, with 3.25 billion reads in total for 8 samples.