GSM7349821: JCML2_scRNA; Homo sapiens; RNA-Seq
Fresh tumor biopsy tissues from an advanced colon cancer patient were transferred to the tube with digestive enzyme from Tumor Dissociation Kit (Cat# 130-095-929, Miltenyi Biotec) and incubated at 37 ℃ for 30 min on a shaker. After digestion, 2% FBS was added to neutralize enzyme lysate, and the tissues were filtered through a 70 μm filter. Subsequently, the samples were centrifuged at 350 × g for 5 min and the supernatants were discarded. To remove red blood cells, the cell pellets were suspended in red blood cell lysis buffer (Beyotime) for 30 s. The solution was then centrifuged at 350 × g for 5 min and resuspended in Dulbecco's phosphate-buffered saline (DPBS; Thermo Fisher). The samples were stained with trypan blue (Solarbio) and the cellular viability was evaluated. Finally, single cells were encapsulated into emulsion droplets using the Chromium Controller (10× Genomics). The scRNA-seq libraries were constructed using the Chromium Single Cell 3' Library, Gel Bead & Multiplex Kit (10x Genomics).