Illumina HiSeq 2500 paired end sequencing; RNA-seq of monocytes from a variety of mouse tissues
RNA-seq of monocytes from a variety of mouse tissues
Single cell suspension of skin, blood, intestine, bone marrow and gingiva were sorted by flow cytometry. Monocytes were identified as CD45+ Lin(CD3ε, TCR-β, CD19, B220, NK1.1, Ter119, Siglec F, Ly6G)- CD11b+ Ly6Chi CD64- MHCII-.  Total RNA from ~700-40,000 FACS sorted monocytes was isolated using the Single Cell RNA Purification Kit (Norgen Biotek Corp). RNA was amplified with the SMART-Seq V3 Ultra Low Input RNA Kit (CloneTech)