GSM5452330: DMSO-treated organoids; Mus musculus; RNA-Seq
Single cell suspensions of the SI organoids were prepared using TrpLE express, stained and epithelial cells were sorted using flow cytometry before proceeding with single cell RNAseq. Libraries were constructed according to 10X Chromium Single Cell 3' Reagent Kits User Guide (v3.1 Chemistry). Briefly, Gel Bead-In-Emulsions (GEMs) were generated with loaded cells in a Chromium Controller. Reverse transcription was performed in GEMs and beads were dissolved. Barcoded cDNA was purified and amplified, then fragmented, A-tailed, ligated to adaptors and sample indeces, and selected for size. Approx 16,000 cells were loaded to achieve approx 10,000 cells final cell counts. Libraries were sequenced by Novogene UK as 150 BP paired-end on NovaSeq600 platform at 300M reads/sample.