title: GSM8689992: WT and GeneX-/-; 0,4,14 days post Cryptococcus infection - CPB; Mus musculus; RNA-Seq
library_construction_protocol: To prepare brain single cell suspensions, tissues within the skull were harvested from mice. Isolated tissues (brain parenchyma including a trace amount of border tissues) were minced and digested in 1X PBS containing 5% heat-inactivated fetal bovine serum, 1mM HEPES, Collagenase D Type IV, and DNase I for 30 minutes at 37°C with constant shaking. Following digestion, tissue was passed through an 25G needle and filtered through a cell strainer. Samples were then resuspended in 38% Percoll and centrifuged at 532 x g for 30 minutes without brake. Following centrifugation, lipid and debris layer was aspirated from the top of the tube and pelleted brain cells were treated with red blood cell lysis. Cells were resuspended in 1X PBS containing 5% heat-inactivated fetal bovine serum, 1mM HEPES. Library was performed according to the manufacturer's instructions using the  Chromium Next GEM Single Cell 3' Kit v3.1 (10x Genomics)