Illumina NovaSeq 6000 paired end sequencing; single cell RNAseq of peripheral CD4+ T cells in the context SARS-CoV-2 infection
single cell RNAseq of peripheral CD4+ T cells in the context SARS-CoV-2 infection
Peripheral (CD14-CD56-CD123-CD19-CD3+CD4+CD45RA-PD-1high) T cells were FACS-sorted using a Sony MA900 in PBS/0.08% FCS. 1X10E5 cells were obtained. Cells were counted and up to 20000 cells were loaded in the 10x Chromium Controller to generate single-cell gel-beads in emulsion. After reverse transcription, gel-beads in emulsion were disrupted. Barcoded complementary DNA was isolated and amplified by PCR. The scRNA-seq libraries were generated using Chromium Next GEM Single Cell V(D)J Reagent Kit v.1.1 with Feature Barcoding (10x Genomics) according to the manufacturer's protocol. Gene expression (mRNA), ADT and VDJ libraries were constructed.