GSM9138363: Non-ITAC; Homo sapiens; RNA-Seq
Fresh tumor tissue samples (≥ 0.2g) were obtained through surgical resection or endoscopic nasal biopsy. The samples were rinsed with 0.9% normal saline and the fat, visible blood vessels and surrounding necrotic areas were removed.Then, the tumor tissue samples were cleaned, minced, and enzymatically digested to generate a cell suspension. Following filtration and centrifugation, red blood cells in the cell suspension were lysed by using a lysis buffer and washed out. The remaining cells were resuspended in RPMI 1640 medium and adjusted to a concentration of 700-1200 cells per microliter. Single-cell suspensions were loaded onto the 10x Chromium chip according to the instructions for the 10x Genomics Chromium Single-Cell 3' kit (V3), with the expectation of capturing 8,000 cells.cDNA amplification and library construction were prepared according to standard protocols. Libraries were sequenced by LC-Bio Technology (Hangzhou, China) on an Illumina NovaSeq 6000 sequencing system (double-end sequencing, 150 bp) at a minimum depth of 20,000 reads per cell.