title: GSM1400987: TR192 Mac virus Positive rep 1; Mus musculus; RNA-Seq
library_construction_protocol: Peritoneal cells from 8-10 mice were harvested by lavage, pooled, and blocked with 10% mouse serum. Cells selected with CD11b MicroBeads (Miltenyi Biotec) were analyzed or sorted after staining with antibodies: CD11b-PerCP-Cy5.5 (clone M1/70, BioLegend), F4/80-APCAlexa Fluor 750 (clone BM8, Invitrogen), CD19-PE-Cy7 (clone 1D3, BD Pharmigen). 3 For analysis cells were also stained with CD206-PE (clone C068C2, BioLegend), CD301-Alexa Fluor 647 (clone ER-MP23, AbD Serotec). Cells were sorted into Trizol Reagent (Invitrogen). 1000 to 2000 cells were obtained from each sort. RNA wasquantified using Qubit RNA assay kits (Invitrogen). All of the RNA from virus-pos cells and 50 ng of RNA from virus-neg cells was prepared for RNA-seq using ScriptSeq v2 RNA-seq library preparation kit (Epicentre). Index Primers (Epicentre) were added and samples underwent Duplex-Specific thermostable nuclease (DSN) (Evrogen) treatment to remove ribosomal RNA. Samples were pooled and sequenced on HiSeq.
identifiers.namespace: GEO
identifiers.id: GSM1400987