GSM6190602: Microglia from naïve mice 3; Mus musculus; RNA-Seq
Immediately after sacrifice mice were perfused with 30 mL of cold 1X PBS. Brains were harvested and put into cold complete RPMI media. For serum, blood from the heart was collected and allowed to clot at 4°C overnight. Brains were minced and homogenized, digested with 0.227 mg/mL collagenase/dispase and 50 U/mL DNase (Roche) at 37°C for 45 minutes. Homogenate was passed through a 70 µm filter (Corning) and washed with cRPMI. Filtered brain homogenate was then resuspended with 20 mL of 40% Percoll and spun at 650 x g for 25 minutes. Myelin was aspirated, samples were washed with cRPMI, and then resuspended in cRPMI. RNA libraries were prepared for sequencing using standard Illumina protocols