GSM7428259: Tuft cells from human colon [HUB-JO-s054]; Homo sapiens; RNA-Seq
Single cell samples were lysed, barcoded and processed according to the CEL-Seq2 technique. Cells were sorted into 384 well plates, containing UMI barcode primers, frozen and incubated at 65°C for 5min to ensure lysis. First and second strand synthesis (Invitrogen) was performed and all wells of a single plate were pooled. Afterinvitrotranscription (Ambion), the amplified RNA was reverse transcribed and amplified for 10-12 cycles with Illumina Truseq primers. Organoids were dissociated into single cells using TrypLE (TrypLE Express, Life Technologies) with 10 μM Rho kinase inhibitor (Abmole) in 37°C and mechanical disruption by pipetting every 5 minutes.