GSM8323119: SW620-Mouse4; Homo sapiens; RNA-Seq
Single cells were dissociated with tumor dissociation kit (Miltenyi Biotec). Then blood cells and mose cells were removed with magnetic beads separation. Finally, live human SW620 cells were counted and diluted in 1x phosphate-buffered saline (PBS) to 700-1200 cells per μL. The cell suspension was loaded in Chromium Next GEM Chip G and sorted in the Chromium Controller from 10X Genomics. The Cell-Gel Beads in Emulsion (GEMs) were then incubated to generate the barcoded cDNA. cDNA was cleaned using Dynabeads and washed, followed by cDNA amplification and SPRIselection. The retrieved cDNA was enzymatically fragmented, end-repaired, poly-A tailed, and ligated. Size selection, adaptor ligation, and amplification were done. Sequencing was done using HiSeq X Ten System (Illumina) according to 10X Genomics specifications. All quality control steps were carried out using Qsep100 System (BIOptic) with High Resolution Cartridge, whereas the concentration was calculated using Qubit 4.0 Fluorometer with Kit High Sensitivity assays.