title: GSM5393313: EGF depletion; Homo sapiens; RNA-Seq
library_construction_protocol: Organoids from control (day21) or EGF depletion (day21) or ADM (day10) condition were incubated with TrypLE select enzyme at 37ºC for 10min. After 10 min, organoids were scooped out using cell scraper (VWR, # 734-2978) and matrigel was disrupted by pipetting. Organoids were again incubated at 37ºC for 10 min followed by TrypLE inactivation using DMEM/F-12 containing 10% FBS medium. Single cell suspension was filtered through 40 µm strainer and dead cells were removed using dead cell removal kit (Miltenyi biotech, #130-090-101) according to manufacturer's instructions. Single cells were resuspended in 0.04% BSA at 1000 cells/µl concentration and used for gel bead emulsion (GEM) generation in 10x chromium instrument. Followed the protocol of 10x Genomics Chromium v3.1 chemistry