title: Illumina NovaSeq 6000 sequencing; Single cell RNA sequencing of murine omental macrophages
design: Single cell RNA sequencing of murine omental macrophages
library_construction_protocol: Mice were injected i.p. with 1x10^6 cells from the mouse epithelial ovarian cancer cell line ID8. Single cell tumor suspensions were prepared from mouse omentum infiltrated with epithelial cancer cells 10 weeks after inoculation by digesting the tissue in RPMI 1640 medium with 1 mg/ml Collagenase II (Sigma Aldrich), 50 µg/ml DNAseI (Roche, Hvidovre, DK) and 0.1% (w/v) BSA for 30 min at 37°C with gentle agitation. Cell suspensions were subsequently passed through 70 µm cell strainer (BD Biosciences, FR) and collected by centrifugation. Macrophages were isolated by flow assisted cell sorting gating on CD11b+, Linneg, F4/80+ and CD64+. Single cell transcriptomes were generated from roughly 4000 cells per mouse using 10x Genomics Chromium Next GEM Single Cell 3' GEM kit v2. Sequencing libraries were generated using 10x Genomics Chromium Next GEM Single Cell 3' GEM kit v2.
attributes.tag: Experimental Factor: individual
attributes.value: 2