scRNA-seq library of wounded (i.e., sham-inoculated) skin from volunteer 481
Single cell suspensions of wound (i.e., sham-inoculated) and pustule punch biopsies were generated with collagenase A, DNase I, and trypsin and processed using the 10X Genomics Single-cell 3' RNA-seq Assay. Each cell suspension were loaded on a multiple-channel micro-fluidics chip of the 10x Genomics Chromium Single Cell Instrument. Single cell gel beads in emulsion containing barcoded oligonucleotides and reverse transcriptase reagents were generated with the Next Gem single cell reagent kit (10X Genomics). Following cell capture and cell lysis, cDNA was synthesized and amplified. At each step, the quality of cDNA and library was assessed by Bioanalyzer. 100-bp reads including cell barcode and UMI sequences were generated on an Illumina NovaSeq 6000. Although volunteers were biopsied on different days, libraries and sequencing of each volunteer's wound-pustule pair were made and processed in the same batch.