title: GSM6213402: Deletion of miR-155 in microglia, biol rep 2; Mus musculus; RNA-Seq
library_name: GSM6213402
library_construction_protocol: The left hemisphere of the mouse brain was homogenized to form a single cell suspension, then resuspended and centrifuged in a 37%/70% Percoll Plus gradient at 800G, 23˚C, for 25 min with an acceleration of 3 and a deceleration of 1. Mononuclear cells were taken from the interface layer. The cells were stained with anti-CD11b PECy7 and CD45 PerCP. After staining cells were washed and sorted using BD FACSAriaTM II. GEMs were generated according to the manufacturer's protocol by combining barcoded Single Cell 3' v3.1 Gel Beads, a Master Mix containing cells, and Partitioning Oil onto Chromium Next GEM Chip G (10X Genomics, USA) with targeted capture of 5000 cells per sample. Reverse transcription immediately followed GEM generation, producing barcoded, full-length cDNA from poly-adenylated mRNA. After incubation, GEMs were broken and pooled fractions of cDNA were recovered. Barcoded, full-length cDNA was amplified via PCR to generate sufficient mass for library construction.