GSM9518226: B1-R493X-16w-o2; Homo sapiens; RNA-Seq
To prepare the cell suspension, organoids were incubated in a solution of Earle's Balanced Salt Solution, 20 units/mL papain, 1mM L-cysteine, 0.5mM EDTA, and 2000 units/mL deoxyribonuclease for 45 minutes at 37ºC. Every 10 minutes, gentle pipetting was used to further break down the organoids. At 45 minutes, media was added to stop the reaction and the tubes were spun down at 300g for 5 minutes. The media was replaced with cold 0.5% BSA in PBS. The samples were filtered once through FACS filter tubes. Additional 0.5% BSA in PBS was used to dilute the samples to reach 1000cells/µL. Collected cells were processed using the Single Cell 3' Gene Expression kit v3 (10x Chromium) according to manufacturer's instructions, and libraries were sequenced an Illumina NovaSeq 6000.