GSM6523061: Control_rep2; Mus musculus; RNA-Seq
Mice were euthanized with CO2 and immediately transcardially perfused with 20 ml 1X PBS. For AD experiments, brains were collected and their meninges and choroid plexus removed then the MACS-sorting protocol was used to isolate microglia as described (Norris et al., 2018). Tissues were homogenized by gently mashing through a 70 um cell strainer.  After centrifugation the cell pellet was resuspended in MACS buffer (Miltenyi Biotec, 130-0910376) to proceed with column purification of microglia using CD11b microbeads (Miltenyi, 130-093-634). We then performed sorting utilizing LS columns and a QuadroMACS magnet (Miltenyi, 130-042-401 and 130-091-051) according to manufacturer's instructions. Column-bound cells were analyzed for purity by flow cytometry and submitted for RNA-seq. RNA libraries were prepared for sequencing by GENEWIZ