GSM5465132: COVID_14_day_5_day_10_PBMC_GEX; Homo sapiens; RNA-Seq
Peripheral blood mononuclear cells were isolated from whole blood by density grandient centrifugation, then washed with Hank's Buffered Saline Solution followed red blood cell lysis using BD Pharm Lyse. Cell were then counted using a Nexcelom Cellometer with acridine orange/propidium iodide for live/dead discrimination. Endotracheal aspirates were extensively diluted with Hank's Buffered Saline Solution, followed by pipetting repeatedly to break up sputum and create single-cell suspensions. Red blood cells were lysed using BD Pharm Lyse. Single cell suspensions of peripheral blood mononuclear cells or endotracheal aspirates were resuspended in 0.04% PBS/BSA and loaded on a 10X Genomics Chip A for use with the 5prime V1 chemsitry kit. In some instances samples were labeled with TotalSeq-C cell hashiing antibodies, and two samples were pooled prior to loading cells for droplet generation. Gene expression and cell hashing libraries were generated following the manufacturer's protocols.