GSM7427939: intest_UT_F_young; Mus musculus; RNA-Seq
Organoids were dissociated into single cell suspensions at 37C using TrypLE, and passed through a 100 um FlowMi cell strainer.  Cells were stained with DAPI and sorted on a Sony SH800S sorter. DAPI-negative viable cells were sorted into a chilled microcentrifuge tube containing PBS. Viability was confirmed to be >75% using a Countess FL II cell counter (Life Technologies #AMQAF1000) before proceeding to single cell library preparation. Single cell libraries were prepared using a 10X Genomics Chromium Controller (cat Chromium Next GEM Single Cell 3' Gene Expression kit (cat #1000268) according to the manufacturer's instructions. Cell suspensions were adjusted to target a yield of 8,000 cells per sample. Single cell 3' digital gene expression with cell barcodes and unique molecular identifiers (UMIs).