title: scRNAseq of mus musculus (strain C57BL/6): adult colon (T3)
design: Single-cell sample preparation was conducted according to the guidelines provided by 10x Genomics. Briefly, a cell suspension (1 ml) from each mouse and condition (triplicates) was pelleted by centrifugation (400 g, 5 minutes). The supernatant was discarded and the cell pellets resuspended in 1X PBS with 0.04% BSA, followed by two washes. After centrifugation (150 g, 3 minutes), cells were resuspended in 500 l 1X PBS with 0.04% BSA followed by gently pipetting 10 to 15 times and automatically counted (Countess automated cell counter, Thermo Fisher Scientific); the viability of cells was assessed by trypan blue staining (0.4%). Subsequently, single-cell Gel beads in Emulsion (GEM) and sequencing libraries were prepared using the 10x Genomics Chromium Controller in conjunction with the single-cell 3' kit (v3). cDNA synthesis, barcoding, and library preparation were performed according to the manufacturers instructions. Libraries were sequenced on an Illumina HiSeq 4000 sequencer (Illumina, San Diego) as paired-end 100 base reads