title: GSM8372404: induced Human Colon Organoid (HCO) in chip co-cultured for 7days, fresh, E4, organoid, scRNAseq; Homo sapiens; RNA-Seq
library_construction_protocol: Dissociated cells and extracted nuclei were respectively suspended in 0.04% bovine serum albumin (BSA) in PBS and 1% BSA in PBS containing 0.2U/µl SUPERase RNase Inhibitor.  Each sample was treated with either ReadyCount Green/Red Viability Stain (Invitrogen) for dissociated cells or YOYO-1 Iodide (ThermoFisher Scientific) for nuclei.  The number of cells/nuclei was measured, and cell viability was checked using the automated cell counter Countess 3 FL (ThermoFisher Scientific). The cell/nuclei suspension was centrifuged at 400 × g for 5 minutes at 4 °C and then resuspended in the BSA solution at an appropriate volume for microfluidic chip loading.  Each sample was loaded per lane of a 10x microfluidic chip device (Chromium Next GEM Chip G).  Generally, for each library, 16,000 cells or nuclei were loaded onto a 10× chromium controller to target the recovery of 10,000 cells or nuclei. Library preparation was carried out using Chromium Single Cell 3′ Reagent Kits (v.3.1), following the manufacturer's instructions.  Libraries were sequenced on a Novaseq system (Illumina) with the aim of achieving a minimum of 30,000 paired-end reads per cell.