title: GSM7795247: Sample_13_Microglia_APOE3_WT; Mus musculus; RNA-Seq
library_construction_protocol: For the isolation of brain microglia, brains were extracted removed from the skull and homogenized to form a single cell suspension, then resuspended and centrifuged in a 30% Percoll Plus (GE Healthcare, 17-5445-02) gradient in HBSS at 800 g, 25°C, for 25 min with an acceleration of 4 and a deceleration of 3. Cells were pelleted and stained with rat APC anti-mouse Fcrls (1:400, clone 4G11, Butovsky Lab). After washing, Ly6c-CD11b+Fcrls+ microglia were sorted using BD FACSAriaTM II (BD Bioscience). Smart-Seq2 libraries were prepared by the Broad Technology Labs and sequenced by the Broad Genomics Platform. cDNA libraries were generated from sorted cells using the Smart-seq2 protocol. RNA sequencing was performed using Illumina NextSeq500 using a High Output v2 kit to generate 2× 25 bp reads.