Single-cell V(D)J-seq of CD3+ and CD19+ lymphocytes from PBMCs of COVID-19 patients
PBMCs were extracted from 5ml of blood by density gradient centrifugation with the help of Lymphoprep. PBMCs were stained with surface markers CD3 FITC and CD19 PB. Cells were sorted in FACS melody. The sorted single cell suspension was converted to single-cell gel beads in the emulsion (GEMs) by using the Chromium Single Cell 5′ Library and Gel Bead kit, and Chromium Single Cell G Chip kit (10x Genomics) according to the manufacturer's protocol. scRNA-seq libraries were constructed using a Single Cell 5′ Library and Gel Bead kit, Single Cell V(D)J Enrichment kit, Human T Cell and a Single Cell V(D)J Enrichment kit, Human B Cell according to manufacturer's protocol. The control, contact and patient sample groups contain 3 individuals each which were tagged using TotalSeq™-C antibodies (C0251, C0252, C0253) respectively.