GSM6857603: WT_aOrg; Mus musculus; RNA-Seq
Organoids were isolated on day 5 after passaging and dissociated into single cells using TrypLE incubation. 20,000 live cells were sorted into 10 mL of ultra clean 0.8% BSA/PBS scRNA libraries were prepared using 10X Genomics v3 Chemistry according to manufacturer's guidelines. Cells were loaded on the 10X Genomics Single Cell Chip for formation of Gel Bead-in-Emulsions (GEMs) and followed by reverse transcriptase. cDNA was amplified with 10 PCR cycles, while sample indexing PCR was performed with 11 cycles. Libraries were diluted to 2 nM in elution buffer, and a final dilution of 1.7 pM was sequenced with Illumina High Output 150 cycles kit on an Illumina NextSeq 500 sequencer.