library_construction_protocol: RNAlater preserved wound samples were stratified into four groups for each wound, Full/Bulk, epidermis, dermis, and subcutaneous. For processing, the previously bisected wound samples (two equal mirror halves of same wound) were thawed from −80°C on ice. Epidermis and Subcutaneous Layers. Epidermis and subcutaneous isolates from the pig wound channel were suspended in 1ml of 4⁰C Invitrogen Trizol Reagent (Fisher Scientific 15-596-026) in a 5ml tissue culture tube on ice. The tissue was homogenized using a Power Gen 35 Tissue Homogenizer employing a medium shark's tooth probe – 6mm diameter (Fisher Scientific) for 20 seconds on ice. The 1ml lysate is transferred to a Andwin Scientific 5Prime Phase Lock Gel – Heavy 2ml tube (Fisher Scientific) and the Phase Lock gel purification is performed according to manufacturer's protocol until the separation centrifugation of the aqueous phase and organic phase is complete. The aqueous phase is transferred to a microcentrifuge tube and the volume is determined. An equal volume of 70% ethanol in nuclease free water is added to the aqueous phase. The ethanol adjusted aqueous phase (with guanidinium salts) is applied directly to a RNeasy Mini kit (Qiagen 74104) purification column and the RNA Clean-up protocol is performed using an on-column DNase treatment with the RNase-Free DNase Set (Qiagen 79254) according to manufacturer's instruction. DNase treated RNA isolates are quality controlled and quantified using a TapeStation 4200 running the RNA ScreenTape assay (Agilent Technologies). RNA Integrity Number (RIN) of ≥8.0 are targeted for proceeding to stranded mRNA library preparation.  Dermis and “Full/Bulk” Layers. Dermis and full tissue isolates (with skin) from the pig wound channel were suspended in 500ul of 4⁰C Invitrogen Trizol Reagent (Fisher Scientific 15-596-026) in a 5ml tissue culture tube on ice. The tissue was homogenized using a Power Gen 35 Tissue Homogenizer employing a medium shark's tooth probe – 6mm diameter (Fisher Scientific) for 20 seconds on ice. The lysate was removed to a new 5ml tissue culture tube on ice. 500ul of additional chilled Trizol Reagent was added to the remaining un-lysed tissue (mainly skin) and homogenized an additional 20 seconds on ice leaving ~10% of the original skin undisrupted. The lysate was pooled with the first lysate for 1ml total. The 1ml lysate is transferred to an Andwin Scientific 5 Prime Phase Lock Gel – Heavy 2ml tube (Fisher Scientific) and the Phase Lock gel purification is performed according to manufacturer's protocol until the separation centrifugation of the aqueous phase and organic phase is complete. The aqueous phase is transferred to a microcentrifuge tube and the volume is determined. An equal volume of 70% ethanol in nuclease free water is added to the aqueous phase. The ethanol adjusted aqueous phase (with guanidinium salts) is applied directly to a RNeasy Mini kit (Qiagen 74104) purification column and the RNA Clean-up protocol is performed using an on-column DNase treatment with the RNase-Free DNase Set (Qiagen 79254) according to manufacturer's instruction. DNase treated RNA isolates are quality controlled and quantified using a TapeStation 4200 running the RNA ScreenTape assay (Agilent Technologies). RNA Integrity Number (RIN) of ≥8.0 are targeted for proceeding to stranded mRNA library preparation. and removed prior to complete thaw so that the tissue was still hard but RNAlater had melted. Samples were immediately transported to the Genomics Core facility in fresh RNAlater (on ice) for RNA processing and isolation. Total RNA was enriched for messenger RNA (mRNA) by oligo dT capture using the NuGEN Universal Plus mRNA-seq + UDI library kit (NuGEN 9144-96). Captured mRNAs were fragmented, reverse transcribed into cDNA and ligated with unique dual indexed (UDI) sequence adapters and PCR amplified to generate indexed libraries. Validation of library preparations was performed using the Agilent Bioanalyzer. Prior to pooling, quantification of the indexed libraries was performed using the Roche LightCycler96 RealTime PCR system.