GSM9571368: B. pertussis infected (4-dpi) 2; Mus musculus; RNA-Seq
Mouse lungs were minced, incubated in digestion buffer composed of collagenase (2.7 mg/mL), hyaluronidase (0.25 mg/mL), and DNase (0.1 mg/mL) for one hour while undergoing gentlemacs lung dissociation at 37C (Miltenyi Biotec), and then 30 seconds of rapid dissaocciation by gentlemacs lung single cell protocol (Miltenyi Biotec). Cell solution was run through a 40 micron filter, pelleted and then resuspended in 1mL of ACK lysing solution for 5 minutes for lysis and removal of red blood cells. Resulting cell mixture was pelleted and resuspended in HBSS buffer w/ 0.04% BSA. Cells were counted by Countess cell counter and aliquoted for downstream processing. Cells were counted using a Thermo Fisher Countess II.  Single cell gene expression libraries were prepared according to manufacturer protocol using the Chromium  Next GEM Single Cell 3' v 3.1 kit.