GSM5741944: C10_pos_D30_Conjunctival_Culture; Homo sapiens; RNA-Seq
Single cell RNA-Seq was performed on cells harvested 24 hours after infection. ALI cultures were washed apically and basally in 1xPBS and dissociated in 100µl 0.025% Trypsin-EDTA (ThermoFisher Scientific) for 15 minutes at 37°C. Trypsin-EDTA was neutralised, and cells were counted on a haemocytometer to get an optimum concentration of 1000 cells per microliter. Suspensions were centrifuged at 400g for 3 minutes and resuspended in 0.04%BSA/PBS solution.  For scRNA-Seq cells were captured and libraries generated using the Chromium Single Cell 3′ Library & Gel Bead Kit, version 3.1 (10x Genomics). scRNA-Seq libraries were sequenced to 50,000 reads per cell on an Illumina NovaSeq 6000.