Spatial RNA-seq library of wounded (i.e., sham-inoculated) skin from volunteer 481
Punch biopsies of wound (i.e., sham-inoculated) and pustule sites were split in half at the bedside and immediately embedded in OCT media and frozen. 10 micron sections of tissue were mounted on the capture areas of a 10X Genomics Visium spatial gene expression slide. Methanol fixation, Hematoxylin & Eosin (H&E) staining, and imaging of tissue were carried out according to the 10x Genomics demonstrated protocol. The tissue was permeabilized for 24 minutes to release the cellular mRNAs for Visium spatial 3' gene expression library construction. Spatially barcoded full-length cDNAs were synthesized and amplified from polyadenylated mRNA captured on the slide. Illumina sequencing libraries were prepared from the amplified cDNA through enzymatic fragmentation, size selection, end-repair, A-tailing, adaptor ligation, and PCR. Each dual-indexed library was quantified and quality assessed with Qubit and Agilent TapeStation.