GSM8956988: Sample_MEPC_DSS; Mus musculus; RNA-Seq
Crypts isolated from mouse intestine were incubated with TrypLE Express (Gibco, 12605010) at 37°C for 60 s. The single cell suspension was passed through a 40-μm filter. The cells were stained with EpCAM (Thermofisher, 14-5791-81) and 7-AAD (BD Biosciences, 559925), and sorted by FACS to enrich 7-AAD-EpCAM+ intestinal epithelial cells. Single-cell RNA sequencing (scRNA-seq) libraries were constructed using the 10 × Genomics single cell 3′ Library and Gel Bead Kit V3 (10 × Genomics, 1000075) following the manufacturer's instructions. The cell suspension (300–600 live cells per mL as determined by Count Star) was loaded onto a Chromium single cell controller (10X Genomics) to generate single-cell gel beads in the emulsion (GEMs). Reverse transcription was performed on a PCR Thermal Cycler (Takara) programed at 53°C for 45 min, followed by 85°C for 5 min to terminate reaction. The cDNA library was generated, amplified, and assessed for quality control using the Agilent 4200. The scRNA-seq was carried out on the Illumina Novaseq6000 sequencer.