GSM6435271: M3A; Mus musculus; RNA-Seq
Mice were sacrificed, excised intestines opened longitudinally, rinsed with cold saline, single cell suspensions prepared, re-suspended in antibody staining buffer, blocked with FcR Blocking Reagent, washed and pelleted, then incubated for 20 minutes at 4°C with, EpCAM-APC and CD45-PerCP. Cells were sorted by FACs on a MoFlo instrument and Epcam positive, CD45 negative total epithelial cells collected. scRNAseq libraries were constructed by the Albert Einstein Genomics Core using the 3' kit protocols of 10X Genomics with approximately 10,000 single FACS collected epithelial cells from each mouse processed on a Chromium Chip B microfluidic apparatus (10X Genomics). Library quality was verified by size analysis of transcripts (BioAnalyzer; Agilent, Santa Clara, CA) and after passing quality control assays, multiple libraries were multiplexed and sequenced by HiSeq PE150 using pair-end sequencing, with a readout length of 150 bases (Novogene; Sacramento, California), and the data assigned to individual cells using the sample indexes.