GSM8941467: HK-374 cells irradiated; Homo sapiens; RNA-Seq
After two weeks, tumors were then harvested and dissociated into single cells using the Miltenyi Brain Tumor Dissociation Kit (#130-095-942), followed by mouse cell depletion (Mouse Cell Depletion Kit, #130-104-694, Miltenyi). Cells from 2-3 mice were pooled, passed through a 40-µm strainer and counted using a hemacytometer. After pelleting by centrifugation at 300 g for 5 minutes at 4°C, the cells were resuspended in the provided Cell Resuspension Buffer. Approximately 50,000 cells were then captured 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.