Single-cell RNA-seq of PBMCs from severe COVID-19 patients

We profiled the single-cell transcriptomes of 13,289 peripheral blood mononuclear cells isolated at three longitudinal stages from two severe COVID-19 patients treated with Tocilizumab. The raw sequencing data can be obtained from the Genome Sequence Archive for Human (GSA-Human) at https://bigd.big.ac.cn/gsa-human/browse/HRA000172 . We generated single-cell transcriptome library following the instructions of single-cell 3’ solution v2 reagent kit (10x Genomics). Briefly, after thawing, washing and countin cells, we loaded the cell suspensions onto a chromium single-cell chip along with patitioning oil, reverse transcription (RT) reagents, and a collection of gel beads that contain 3,500,000 unique 10X Barcodes. After generation of single-cell gel bead-in-emulsions (GEMs), RT was performed using a C1000 TouchTM Thermal Cycler (Bio-Rad). The amplified cDNA were purified with SPRIselect beads (Beckman Coulter).