title: RNA-SEQ of Human skin: diabetic foot ulcers
design: Tissue from wound margins was collected during the standard of care and debridement, and preserved in RNALater (Life Tech). Tissue samples were homogenized, total RNA was extracted with the Quick-RNA Mag Bead kit (Zymo Research), and the integrity of the RNA was evaluated using an Agilent Tape Station (Agilent). For sequencing, indexed libraries were constructed using the SMARTer Stranded Total RNA-Seq Kit v3 (Takara Bio). Both the quantity and quality of these libraries were appraised by a Qubit fluorometer and an Agilent 2100 bioanalyzer. Molar concentrations of the libraries were confirmed by qPCR prior to pooling. The sequencing process was conducted on the Illumina NovaSeq 6000 platform, utilizing PE150 chemistry (Illumina).