GSM7779365: TPF-resistant Fadu cell line, treated with TPF + Cinobufotalin, scRNAseq; Homo sapiens; RNA-Seq
The cell suspension was centrifuged at 300r at 4℃ for 5min. Discard the supernatant and add 1mL of re-suspension to the centrifuge tube to re-suspend the cells, add 3mL of pre-cooled erythrocytes lysis liquid in tube (Solarbio), blowing and suction evenly. Incubate at 4℃ for 5-10min, centrifuge at 300r at 4℃ for 5min immediately after completion and remove the supernatant and add 1mL of re-suspension( Precooled 1X PBS) to fully re-suspend the cells. Test with cell counter（Countstar Rigel S2）, adjust the concentration according to the test result, the target concentration is 700-1200 cell/µL, the Cell activity is greater than 85%, and the agglomeration is less than 15%. Once the final cell concentration and activity were reached, the cells were placed on ice and 10x Genomics single cell transcriptome chip on-board experiment was carried out within 30 minutes. Single cell RNA-seq libraries were performed with Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 on the Chromium Controller(10× Genomics).The quality of libraries was assessed by Qubit 4.0 and the Agilent 2100. Sequencing was performed on the Illumina NovaSeq 6000 with a sequencing depth of at least 50,000 reads per cell and 150 bp (PE150) paired-end reads.