title: GSM6249885: Mock 3dpi Set 2; Homo sapiens; RNA-Seq
library_construction_protocol: Mock and KSHV infected tissues were harvested at 3 days and 6 days post infection. Tissue inserts were removed from the 6-well plate and placed into a new 6-well plate with enough PBS to coat the bottom. The inside of the insert was then washed with PBS to collect the supernatant. The tissues where then carefully separated from the insert membrane and chopped into small pieces in DMEM. The cells are then incubated for 40 minutes in 2 mL of 1 mg/mL collagenase and dispase (Roche Cat. 10269638001) in a shaking incubator set to 37ºC and 250 rpm. The collagenase and dispase are then inactivated by adding 2 mL of DMEM and 10% FBS. The digested cells are then passed through a 0.45 um cell strainer and then washed with cold PBS with 0.04% BSA (ThermoFisher Cat. AM2618). The filtered cells are also passed through a flowmi cell strainer. Cells are counted with Invitrogen Countess II (ThermoFisher Cat. A27977). Single Cell Libraries were prepared with the 10X Genomics Single Cell Gene Expression v2 kit (Set 1 and 2) or v3 kit (Set3). The Chromium Single Cell 3ʹ Reagent Kits v3 User Guide Rev A protocol was used for sample preparation and library generation.