GSM8080508: Pooled WT colon scRNA-seq; Mus musculus; RNA-Seq
Single cell suspensions of colons from 9-month-old littermate WT and 5XFAD female mice were generated using a lamina propria dissociation kit (Miltenyi Biotec, 130-097-410) with the gentleMACS Dissociator (Miltenyi Biotec, 130-093-235). Cells were stained with LIVE/DEAD Fixable Blue cell stain (Thermofisher, L23105) for viability assessment and blocked with FcBlock with CD16/32 (Biolegend, 101320). Cells were then stained with anti-mouse CD45 (Biolegend, 30-F11). Live CD45+ cells were sorted using the FACS Aria sorter at the Buck Institute for Aging Research flow cytometry core facility and collected into RPMI-1640 (Wisent) with 10% FBS (Wisent). Samples were processed utilizing a Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1. The resulting libraries were sequenced at UC Davis Genome Center Bioinformatics Core using Illumina NovaSeq 6000 sequencing.